Looking for Cell Counting Kit-8 protocol?

The Cell Counting Kit-8 is a colorimetric assay kit used to measure cell proliferation and cytotoxicity. It is a ready-to-use solution that can be added directly to the cell media for fast, high-throughput screening without a solubilization process obtaining highly reproducible and accurate results.

Cell Counting Kit-8 can be utilized to do cell number counting, cell prolifiration or cytotoxicity assay based on your requirments.  So it make a tiny protocol deferences when you apply different assay, while the majority of  operation protocol are the same. However, Some equipment and materials should be prepared before assay: 1). Plate reader (450 nm filter), 2). 96-well plate, 3). 10 µl, 100-200 µl and multi-channel pipettes.

 



More detailed manipulation procedures for Cell number counting and Cell proliferation and cytotoxicity sssays from Abbkine CCK-8 Kit are listed below for your references,

Cell Number Determination Protocol

1) Inoculate cell suspension (100 µl/well) in a 96-well plate. Also prepare wells that contain known numbers of viable cells (to create a calibration curve in step 5). Pre-incubate the plate in a humidified incubator (e.g., at 37 °C, 5% CO2).

2) Thaw the CCK-8 on the bench top or in a water bath at 37 °C if it is frozen. Note: It takes about 30 minutes on the bench top at 25 °C or 5 minutes in a water bath at 37 °C.

3) Add 10 µl of the CCK-8 solution to each well of the plate. Note: Be careful not to introduce bubbles to the wells, since they interfere with the O.D. reading.

4) Incubate the plate for 1-4 hours in the incubator.

5) Measure the absorbance at 450 nm using a microplate reader. Prepare a calibration curve using the data obtained from the wells that contain known numbers of viable cells.

Note: To measure the absorbance later, add 10 µl of 1% w/v SDS to each well, cover the plate and store it with protection from light at room temperature. No absorbance change should be observed for 48 hours.

Cell Proliferation and Cytotoxicity Assay Protocol

1) Dispense 100 µl of cell suspension (5000 cells/ well) in a 96-well plate.

2) Pre-incubate the plate for 24 hours in a humidified incubator (e.g., at 37 °C, 5% CO2).

3) Add 10 µl of various concentrations of toxicant into the culture media in the plate.

4) Incubate the plate for an appropriate length of time (e.g., 6, 12, 24 or 48 hours) in the incubator.

5) Thaw the CCK-8 on the bench top or in a water bath at 37 °C if it is frozen. Note: It takes about 30 minutes on the bench top at 25 °C or 5 minutes in the water bath at 37 °C.

6) Add 10 µl of CCK-8 solution to each well of the plate. Be careful not to introduce bubbles to the wells, since they interfere with the O.D. reading.

7) Incubate the plate for 1-4 hours in the incubator. Measure the absorbance at 450 nm using a microplate reader.

Note: To measure the absorbance later, add 10 µl of 1% w/v SDS to each well, cover the plate and store it with protection from light at room temperature. No absorbance change should be observed for 48 hours.

Be attention to conditions or chemicals that affect dehydrogenase activity in viable cells may cause discrepancy between the actual viable cell number and the cell number determined using the Abbkine CCK-8 assay.

Anti-Nano-Tag9 Monoclonal Antibody(11T3) Review

The Nano-tag is a new streptavidin-binding peptide for both the purification and the detection of Nano-tagged proteins. This peptide possesses nanomolar-affinity for streptavidin and therefore is termed Nano-tag. The nano-tags have two types, Nano-tag15 (MDVEAWLGARVPLVET) and Nano-tag9 (MDVEAWLGAR), which bind to streptavidin with dissociation constants of 4 nM and 17 nM, respectively. The Nano-tag(15) is 15 amino acids long and binds to streptavidin with a dissociation constant of 4 nM and the Nano-tag(9) is a 9-mer peptide with a dissociation constant of 17 nM.

Anti-Nano-Tag9 Monoclonal Antibody(11T3) was affinity-purified from mouse ascites by affinity-chromatography using specific immunogen. This antibody has been tested with WB. And Abbkine suggested starting dilutions are as follows: WB 1:2000-5000.

Well-characterized antibodies for epitope tags consisting of short sequences are widely used in the study of protein expression in various systems. We find the quality suitable for our experiments. This product is what I want.

 

Choose right VSV-G Tag Antibody?

Epitope tags include VSV-G tag, Myc-tag, HA-tag and NE-tag. These tags are particularly useful for western blotting, immunofluorescence and immunoprecipitation experiments, although they also find use in antibody purification.

If you’re working in the life sciences with VSV-G tag in your vector or fusion proteins, chances are you’re going to be using right VSV-G antibodies in your research. Here we've set out some top tips to get you started in making a good and suitable VSV-G tag antibody buying decision.

Is this VSV-G antibody suitable for your application?

It's “impossible” to predict which antibodies will work and which will not.  Just because an antibody has been successfully used in one application does not mean it will work for all applications. That means excellent WB and IHC results do not tell you whether the antibody is the right choice for a so-called ChIP-seq experiment, in which chromatin immunoprecipitation is followed by second-generation sequencing.

You must choose application-specific validation of VSV-G antibody you use. However, due to the price and performance, researchers prefer the antibody with more validated applications, especially these kinds of epitope antibodies.

Decision on monoclonal or polyclonal VSV-G tag antibody

Our advice is you should go with the most validated antibody, but if you are lucky enough to have two well validated antibodies to chose between, then polyclonal antibodies tend to be cheaper because they are raised against multiple epitopes on your protein and are therefore less specific. Monoclonal are more expensive and take longer to produce, however they are specific to a single epitope. Monoclonal antibodies are most useful if you are trying to identify specific isoforms or region of a full-length protein, especially in tag fusion detection.

[caption id="attachment_14" align="aligncenter" width="384"] Immunofluorescence staining of VSV-G fusion protein with VSV-G monoclonal antibody (Clone #14D2) in 293 cells with red and counterstained with DAPI.[/caption]

A manufacturer may license their antibody to other companies, so do not buy three antibodies to test without making sure they are actually different. With common monoclonals that are supplied by many companies you might also consider shopping around for the best supplier of a clone.

Finally, remember you should feel free to contact the company supplier for advice any time. The suppliers really should be helpful, many pride themselves on their customer service, and if they are not you can remember that for next time you buy an antibody.

If you are looking for a detection tool to quantify or localize your VSV-G fusion proteins, we highly recommend you to try our featured monoclonal VSV-G Antibody risk free. This antibody with clone number 14D2 can specifically recognizes native and denatured forms of VSV-G fusion proteins, with which can meet your any requirements for Western Blot, Immunofluorescence and Immunoprecipitation assays.

Preparing Cell Counting Kit-8 Solution ?

Cell Counting Kit-8 (CCK-8) solution in kit assign very convenient assays by utilizing highlywater-relaxed tetrazolium sailor, WST-8 to produce a water-dissolvable formazan Dyeupon decrease in the person of an electron porter, as shown in below.  WST-8 is educed by dehydrogenases in cells to give a yellowcolored roduct (formazan), which is soluble in the membrane civilization medium. The amount of the formazan dye generated by the nimbleness of dehydrogenases in cells is soon relative to the number of ignited cells.

"The CCK-8 Kit has been optimized for use." Said Brooke Kilyanek, Technical Manager of Abbkine Scientici. "If you want to buy WST-8 and 1-methoxy-PMS, and then mix the two regents  together to Cell counting kit-8 solution for cell proliferation assays yourselves. This is also certainly an option, but the best protocol to use is ultimately up to the researcher. "

Cell Counting Kit-8 is a one-deadman solution, and no mixing of components is required, which is totally differently from other commercial player in the market, such as WST-8 Kit. WST-8 Kit is normal made from two solution components. One component is an electron carrier for NADH-tetrazolium, which is 1-methoxy-PMS in most cases. And another component is WST-8 solution with proper reaction buffer.  While Cell Counting Kit-8 featured only one optimized solution component, which is nonradioactive, allot sensitive colorimetric assays for the purpose of the reckon of viable cells proliferation and cytotoxicity risk.

Since the CCK-8 solution is very stable and it has weak cytotoxicity, a longer incubation, such as 24 to 48 hours, is possible. We highly suggest you purchase bulk size of Cell Counting Kit-8 if you're willing to prepare CCK-8 solutions yourself. Since this will not only saving your time and cost, but also ensuring you reliability of cell research. Believe me, high quanlity branded CCK-8 allows you archieve colorimetric assays for the determination of the number of viable cells in the proliferation and cytotoxicity assays with high performance and high throughput.

How to Choose right Cell Counting Kit-8 product

Choosing a cell viability or cytotoxicity assay from among the many different options available can be a challenging task. Picking the best assay format to suit particular needs requires an understanding of what each assay is measuring as an endpoint, of how the measurement correlates with cell viability, and of what the limitations of the assay chemistries are.

Cell Counting Kit-8 allows sensitive colorimetric assays for the determination of the number of viable cells in the proliferation and cytotoxicity assays. CCK-8 highly water-soluble tetrazolium salt, WST-8, is reduced by dehydrogenase activities in cells to give a yellow-color formazan dye, which is soluble in the tissue culture media. Cell Counting Kit-8 is a one-bottle solution; no premixing of components is required. Cell Counting Kit-8, being nonradioactive, allow sensitive colorimetric assays for the determination of the number of viable cells n cell proliferation and cytotoxicity assays.

PurKine™ Anti-Flag Resin 4FF Released

FLAG-tag,  is a polypeptide tag that can be added to a protein using recombinant DNA technology, having the sequence DYKDDDDK. It has been used for studying proteins in living cells and for protein purification by affinity chromatography. It has been used to separate recombinant, overexpressed protein from wild-type protein expressed by the host organism. Because of the small size of the FLAG peptide tag, it is not likely to obscure other epitopes, domains, or alter function, secretion, or transport of the fusion protein. The peptide sequence of the FLAG-tag from the N-terminus to the C-terminus is: DYKDDDDK (1012 Da).

PurKine™ Anti-Flag Resin 4FF has been designed for affinity purification of Flag fusion proteins.The Resin consists of 90μm beads of cross-linked 4% agarose, to which mouse monoclonal antibody against DYKDDDDK tag has been coupled. Tests confirm that performance equals or exceeds popular Anti-Flag resins from other suppliers, and no decrease in performance occurs after at least five repeated uses. In addition, its high flow properties make it excellent for scaling-up.

More than 1mg DYKDDDDK-tagged protein can be captured by per mL of PurKine™ Anti-Flag Resin 4FF. The maximum tolerance pressure of the resin is 0.3MPa, 3bar, which is excellent for industry production. In addition,  Anti-Flag Tag Mouse Monoclonal Antibody (1B10) is a good option to detect the Flag fusion protein that is affinity purified.

Anti-AmCyan Monoclonal Antibody (8T2) Review

AmCyan is one of the existing violet fluorochromes for use in flow cytometers equipped with a violet (405 nm) laser. It is also widely used as a label in fluorescent spectroscopy. AmCyan was originally derived from the fluorescent protein amFP486. AmFP486 was cloned from the sea anemone Anemonia majano (GenBank accession number AF168421), and belongs to the family of fluorescent proteins (FPs) isolated from coral reef organisms; green fluorescent protein (GFP), derived from the jellyfish Aequorea victoria, is its most famous representative.AmCyan has been
adapted from the corresponding full length cDNA for higher solubility, brighter emission, and rapid chromophore maturation. AmCyan1 has been human-codon optimized for enhanced translation in mammalian cells.

Anti-AmCyan Monoclonal Antibody (8T2) was affinity-purified from mouse ascites by affinity-chromatography using specific immunogen. This antibody has been tested with WB. And Abbkine suggested starting dilutions are as follows: WB 1:5000.

AmCyan is a very good blue fluorescent reporter protein, and useful in two-color analyses with DsRed2, DsRed-Express, or AsRed2, and in three-color analyses with ZsYellow and HcRed. I've tried Anti-AmCyan Monoclonal Antibody (8T2) with excellent results. I am a loyal fan of Abbkine！

PurKine™ Antibody Purification NHS-Activated Resin 4FF Released

NHS-Activated Resin contains N-hydroxysuccinimide (NHS) functional groups. NHS coupling forms a chemically stable amide bone with ligands containing primary amino groups. NHS-Activated Resin can be used to prepare affinity purification procedures which can isolate specific substances from complex mixtures, often achieving very high purity in a single step.

PurKine™ Antibody Purification NHS-Activated Resin 4FF is a pre-activated agarose matrix that increases the choice of coupling chemistries available. It is suitable for coupling amino-containing smaller proteins and peptides. NHS-Activated Beads 4FF is crosslinked, 4% beaded agarose resin that contains N-hydroxysuccinimide (NHS) functional groups.

More than 10mg human lgG can be captured by per mL of NHS-Activated Resin 4FF. Tests confirm that performance equals or exceeds popularNHS-Activated Resin from other suppliers, and no decrease in performance occurs after at least five repeated uses. In addition, its high flow properties make it excellent for scaling-up.

Test Post from Cell Counting Kit-8 for your Cell Proliferation and Cytotoxicity Assay

Test Post from Cell Counting Kit-8 for your Cell Proliferation and Cytotoxicity Assay
http://www.cellcountingkit8.com

Anti-EYFP-Tag Monoclonal Antibody(10T3) Review

Yellow Fluorescent Protein (YFP) is a genetic mutant of green fluorescent protein (GFP) originally derived from the jellyfish Aequorea victoria. Its excitation peak is 514 nm and its emission peak is 527 nm. YFP is a useful tool in cell and molecular biology thanks to its properties useful for fluorescence microscopy. Typically, yellow FPs serve as the acceptor for genetically-encoded FRET sensors of which the most likely donor FP is mCFP (monomeric cyan FP).

Anti-EYFP-Tag Monoclonal Antibody(10T3) was affinity-purified from mouse ascites by affinity-chromatography using specific immunogen. This antibody has been tested with WB. And Abbkine suggested starting dilutions are as follows: WB 1:5000.

Like green fluorescent protein (GFP), YFP is a useful tool in cell and molecular biology, usually explored using fluorescence microscopy. I am happy to choose Anti-EYFP-Tag Monoclonal Antibody(10T3). The pictures are beautiful!

Choose right HA Tag Antibody?

Epitope tags include HA-tag, VSV-G tag, Myc-tag and NE-tag. These tags are particularly useful for western blotting, immunofluorescence and immunoprecipitation experiments, although they also find use in antibody purification.
If you’re working in the life sciences with HA tag in your vector or fusion proteins, chances are you’re going to be using right HA antibodies in your research. Here we've set out some top tips to get you started in making a good and suitable HA tag antibody buying decision.Is this HA antibody suitable for your application?

It's “impossible” to predict which antibodies will work and which will not.  Just because an antibody has been successfully used in one application does not mean it will work for all applications. That means excellent WB and IHC results do not tell you whether the antibody is the right choice for a so-called ChIP-seq experiment, in which chromatin immunoprecipitation is followed by second-generation sequencing.

You must choose application-specific validation of HA antibody you use. However, due to the price and performance, researchers prefer the antibody with more validated applications, especially these kinds of epitope antibodies.

Decision on monoclonal or polyclonal HA tag antibody

Our advice is you should go with the most validated antibody, but if you are lucky enough to have two well validated antibodies to choose between, then polyclonal antibodies tend to be cheaper because they are raised against multiple epitopes on your protein and are therefore less specific. Monoclonal are more expensive and take longer to produce, however they are specific to a single epitope. Monoclonal antibodies are most useful if you are trying to identify specific isoforms or region of a full-length protein, especially in tag fusion detection.

[caption id="attachment_26" align="aligncenter" width="489"] IP (1:200) - WB (1:5,000) analysis of HA fusion protein expression in 293 cells. Untransfected 293 cell lysate (lane A), transfected 293 cell lysate with HA-tag protein (lane B); IP untransfected 293 cell lysate with Anti HA tag mAb (lane C); IP transfected 293 cell lysate with normal Mouse IgG (lane D), or with Anti HA tag mAb (lane E), and IP transfected 293 without both normal Mouse IgG and HA tag mAb (lane F).[/caption]

A manufacturer may license their antibody to other companies, so do not buy three antibodies to test without making sure they are actually different. With common monoclonals that are supplied by many companies you might also consider shopping around for the best supplier of a clone.

Finally, remember you should feel free to contact the company supplier for advice any time. The suppliers really should be helpful, many pride themselves on their customer service, and if they are not you can remember that for next time you buy an antibody.

If you are looking for a detection tool to quantify or localize your VSV-G fusion proteins, we highly recommend you to try our featured monoclonal HA Antibody risk free. This antibody with clone number 4F6 can specifically recognizes native and denatured forms of VSV-G fusion proteins, with which can meet your any requirements for Western Blot, Immunofluorescence and Immunoprecipitation assays.

PurKine™ Antibody Purification SulfoLink Resin Released

SulfoLink Resin  is a simple and efficient affinity chromatography medium. It allows covalent immobilization of sulfhydryl containing peptides, protein and other ligands to the medium agarose support for use in affinity purification procedures. The medium reacts specifically with free sulfhydrys to form a stable thioether linkage.


PurKine™ Antibody Purification SulfoLink Resin consists of average particle sizeof 90µm beads of 4% agarose beads with iodoacetyl groups, any molecule that contains free (reduced) sulfhydryl groups can be immobilized on SulfoLink Resin. User can get high valence of antibody by the resin, it is the indispensable affinity chromatography medium of polyclonal antibody purification.

PurKine™ Antibody Purification SulfoLink Resin is ideal for conjugating sulfhydryl containing peptide for subsequent antibody purification. It reacts specifically and efficiently with the exposed sulfhydryls (-SH).  One ml SulfoLink Resin can capture at least 3mg IgG. The resin is the best choice for custom-made affinity for purification of antibodies, antigens and other molecules of interest.

Anti-Strep-Tag II Monoclonal Antibody(8C12) Review

Strep-tag II antibody detects amino acid sequence specific for Streptavidin. Streptavidin is a tetrameric protein purified from Streptomyces avidinii. It has wide use in numerous molecular biological protocols dues to its strong affinity for biotin. The original Strep-tag is a nine amino acid peptide with high specificity and affinity towards streptavidin. The addition of this tag to the C-
terminus of recombinant proteins allowed the simple purification of protein by use of affinity columns. It was re-engineered (and re-named Strep-tag II) to allow it to also placed at the N-terminus of recombinant proteins. Anti-Strep-tag II antibodies are ideal for investigators involved in GFP and Epitopes research.

Anti-Strep-Tag II Monoclonal Antibody(8C12) was affinity-purified from mouse ascites by affinity-chromatography using specific immunogen. This antibody has been routinely tested with WB applications. And Abbkine suggested starting dilutions are as follows: WB 1:2000-5000.

Streptavidin is a tetrameric protein purified from Streptomyces avidinii. It has wide use in numerous molecular biological protocols dues to its strong affinity for biotin. It is responsible to say that Anti-Strep-Tag II Monoclonal Antibody(8C12) has a reliable quality.

 

How to order?

How to place an order?

We accept orders by the following methods:

• Online : To place an order online, search for the product(s) you would like and click on the "Add to cart" button to add them to your shopping cart.


• Offline: Orders can be sent by directly email to service@abbkine.com or you can contact our distributors in your country.

Order procedure

• If you are ready to place an order, please contact service@abbkine.com or customer service of your region, then you will be informed of the Item Number, Size, Delivery date, and Price in detail.


• After you place an order, Pro-forma invoice(p/I) would be sent to you by email for you to check and confirm.


• After you confirm the P/I, please pay the bill. On receipt of the payment, we would prepare the cargo immediately. Any changes of your order information, please inform our customer service for modification.


• The goods will be delivered within three to five days on receipt of payment. The time of special detection item may be longer. We will contact you in advance if having any changes of delivery date or shipment to ensure the experiment will be operated as schedule.

How to pay?

• We accept bank transfer, Paypal, credit card as methods of payment.


• If you place an order online, extra $80 transportation expense will be paid. Under current condition, we recommend you choose offline bank transfer or Paypal transfer to account@abbkine.com.

About shipping

• We recommend you choose the following express company (FedEx, UPS, DHL, TNT, EMS) in which you have an account. You will be informed of the track information, the Shipment Number will be provided to you by email once having delivered the goods, so you can also track the order through the website of the express company. In this case, you only need pay cargo fee to us. You will responsible for transportation expense and local customer clearance fee.


•  On receipt of the goods, please check whether the reagent is complete. If the reagent is damaged, please remember to take pictures of the damaged product and contact us so that we can verify the information and solve the problem for you as soon as possible.

For further details, please email us at service@abbkine.com. We'll contact you once we receive your email.

AmCyan Sequence

AmCyan is a cyan fluorescent protein designed from Clontech (Now part of Takara). The living color protein was isolated from the coral reef organism Anemonia majano. Cyan fluorescent proteins such as AmCyan1 are ideal for simultaneously detection of two or more events in the same cell or cell population, because their excitation and emission spectra are distinct from other fluorescent proteins.

The AmCyan fluorescent protein sequence has been optimized for translation in mammalian cells with high solubility, bright emission, and rapid chromophore maturation. AmCyan1 can be used as a molecular tag or as a reporter to visualize, track, and quantify cellular processes including protein synthesis and turnover, protein translocation, gene induction, and cell lineage.

AmCyan DNA sequence in pAmCyan vector 

pAmCyan is a pUC19-derived prokaryotic expression vector, which encodes a variant of wildtype Anemonia majano cyan fluorescent protein (AmCyan 1) that has been engineered for brighter fluorescence. Two amino acid substitutions (Asn-34 to Ser; Lys-68 to Met) have been made to enhance the emission characteristics of AmCyan (excitation maximum = 458 nm; emission maximum = 489 nm).



As shows above, the sequence of Anemonia majano Cyan fluorescent protein gene is from site 282 to 978. The fist part is Kozak consensus translation initiation site from 282 to 292, and then start codon (ATG) is from 289-291 until stop codon from 976 to 978. You may check for detailed pAmCyan vector sequence and map information.

AmCyan protein sequence

DNA sequence of AmCyan ORF is 690bp, which code total 229 amino acid of enhanced variant AmCyan proteins. Two amino acid mutations are designed in Asn-34 to Ser mutation (A->G) at 389 site, and Lys-68 to Met mutation (A->T; A->G) at 491 and 429 sites, separately.

MALSNKFIGDDMKMTYHMDGCVNGHYFTVKGEGSGKPYEGTQTSTFKVTMANGGPLAFSFDILSTVFMYGNRCFTAYPTSMPDYFKQAFPDGMSYERTFTYEDGGVATASWEISLKGNCFEHKSTFHGVNFPADGPVMAKMTTGWDPSFEKMTVCDGILKGDVTAFLMLQGGGNYRCQFHTSYKTKKPVTMPPNHAVEHRIARTDLDKGGNSVQLTEHAVAHITSVVPF

We offer both monoclonal AmCyan Antibody and polyclonal AmCyan Antibody which recognizes native and denatured forms of AmCyan1 fluorescent protein, as well as fusion proteins containing AmCyan1.

PurKine™ Protein AT (alkaline tolerate) Resin 4FF Released

Protein At is an alkali-resistant protein A (Alkaline tolerate) based on the native protein A though biotechnology mutation. Protein At Resin has been immobilized for the capture of monoclonal antibodies at process scale. Protein At Resin provides greater stability than conventional protein A-based resin in  alkaline conditions used in cleaning-in-place (CIP) . The enhanced alkali stability of protein At Resin improves process economy and product quality.

PurKine™ Protein AT (alkaline tolerate) Resin 4FF consists of 90μm beads of cross-linked 4% agarose, to which Recombinant protein A (alkaline tolerate) has been coupled. The resin withstands rigorous CIP and sanitization procedures with 0.1 to 0.5 M NaOH. The resin load is almost unchanged after hundreds of CIP. At mean time, it's a robust, high-flow resin that ensures excellent pressure/flow properties, low non-specific binding , and an oriented ligand coupling for optimal binding capacity.

The binding capacity of PurKine™ Protein AT (alkaline tolerate) Resin 4FF is  more than 40mg human lgG for per ml of resin. Tests confirm the resin has an increased stability towards proteases, leading to lower ligand leakage. For large-scale industry production, Protein AT (alkaline tolerate) Resin 4FF maybe your best choice as CIP can be performed with cost-effective reagents such as sodium hydroxide instead of expensive and hazardous cleaning agents such as Gua-HCl.

Anti-TAP Tag Monoclonal Antibody(4H2) Review

Tandem affinity purification (TAP) is a purification technique for studying protein–protein interactions. It involves creating a fusion protein with a designed piece, the TAP tag, on the end.The TAP tag historically consists of a calmodulin binding peptide (CPB), a tobacco etch virus (TEV) protease cleavage site, and Protein A. However, additional tag combinations have been used with the TAP method including the combination of FLAG tags and HA tags.The TAP method permits the identification of proteins interacting with a particular target protein without any prior knowledge about the function, activity, or composition of the interacting proteins. This tag is also known as the C-terminal TAP tag because an N-terminal version is also available. However, the method to be described assumes the use of a C-terminal tag, although the principle behind the method is still the same.

Anti-TAP Tag Monoclonal Antibody(4H2) was affinity-purified from mouse ascites by affinity-chromatography using specific immunogen. This antibody has been tested with WB. And Abbkine suggested starting dilutions are as follows: WB 1:5000-10000.

The TAP (Tandem Affinity Purification) method is an affinity purification method for the isolation of TAP-tagged proteins along with associated proteins. To my knowledge, Abbkine is a great option when you want to find a cost-effective product. It isworth recommending.

International Distributors Wanted

Abbkine Scientific Co., Ltd. was founded by a number of scientists and marketing experts in the field of life science in California in 2012. With growing demands from Asia Pacific, it move its headquarters to China. Abbkine started featured and exclusive products from protein detection tools, and it's devoted to developing and providing the most innovative solutions for global customers covering all kinds of antibodies, biochemicals, proteins, assay kits.

As the expansion of Abbkine’s business, in order to provide integrated portfolio and on-site technical support to our clients all around the world, we are looking for distributors reselling products and providing  services to the local customers faster and better.

By cooperating with Abbkine, you can not only benefit a lot from a good price discount and marketing revenue margin, but also from:

• Growing comprehensive life science research portfolio with aggressive market competitiveness.


• Active and professional support to delight your clients with 100% satisfaction guarantee.


• Designated business development specialist and technical engineers for partner support.

For us, every distributor is unique and our cooperation could be flexible and based on the case-by-case approach. So, we are looking forward to discussing with you about any possibilities of cooperation. Abbkine is focusing on providing innovative, high quality and cost-effective life science research and diagnosis products, and professional and quick-response service to the customers all around the world so that they can enjoy the comfort and convenience of biology technology. We understand that it is not an easy work to introduce a new product to a new market. It requires continuous efforts and investment from both sides. Therefore, we are looking for setting up long-term cooperation with those who would like to grow with Abbkine based on mutual benefit principles.

Key Benefits of being a Abbkine Distributor

• FINANCIAL BENEFITS
  
  • Generous price margin
  
  
  • Support with low cost
  
  
  • Flexible pricing policy
  
  


• MARKETING AND SALES SUPPORT
  
  • Up-to-date enterprise website
  
  
  • Proper use of Abbkine Logo
  
  
  • Expo/Emarketing Support
  
  
  • Gifts with Abbkine Logo
  
  


• TECHNICAL AND TRAINING SUPPORT
  
  • Email and phone support
  
  
  • Demo program support
  
  
  • Cytology and Protein solution consultancy
  
  

HOW TO BECOME ABBKINE DISTRIBUTOR?

• Step 1: Send us with your company profile and letter of motivation.


• Step 2: Discuss the cooperation module and agreement.


• Step 3: Sign the distributor agreement.

Abbkine has established business and research partnership with Merck-Millipore, Bio-Techne, Abnova, and Jackson Immunoreagents. Abbkine warmly welcomes companies from all over the world to cooperate with us to provide Life science products and technical services to different local market. If you are interested in our product range and you believe we could be the right partner for your business, please do not hesitate to contact us at marketing@abbkine.com. To learn more about Abbkine, please visit www.abbkine.com.

Versatile PurKine™ Protein L Resin 4FF

Protein L is a bacterial surface protein isolated from Peptostreptoccocus magnus, and recombinant protein L is produced in E.coli. Unlike Protein A and Protein G, which bind primarily through Fc regions of immunoglobilins, Protein L binds Igs through interactions with the light kappa chain of all classes of immunoglobulin such as IgG, IgM, IgA, IgE, IgD, and Fab fragments. It also binds to single-chain antibody fragments (scFv).

PurKine™ Protein L Resin 4FF  has been designed  to purify mammalian IgG that contain specific kappa light chains from serum, ascites fluid, cell culture supernantant and other antibody samples.The Resin consists of 90μm beads of cross-linked 4% agarose, to which Recombinant protein L has been coupled. More than 15mg human lgG can be captured by  per mL of this resin. No decrease in performance occurs after at least five repeated uses. In addition, its high flow properties make it excellent for scaling-up.

In a word, PurKine™ Protein L Resin 4FF is a versatile affinity resin which can be apply in purifying monoclonal and polyclonal antibodies which bind poorly to Protein A or Protein G, monoclonal antibodies from culture supernatants supplemented with bovine serum ( it does not bind bovine immunoglobilins), immunoprecipitation and co-immunoprecipitation assays, ScFv and Fab fragments containing kappa light chains.

How to Choose good Amcyan antibody

One of the most frustrating things in the lab is spending good money on an antibody only to find out it doesn’t work well in your application. You can then either spend weeks trying to make it work, abandon your work altogether or humbly ‘fess up that you made a bad purchase and beg for a second chance. In an effort to limit your groveling, we have made a list of tips for choosing a good Amcyan antibody for Amcyan epitope or Amcyan fusion proteins.

Pay attention to application

The best case scenario is to find a Cyman tag antibody that has already been used in the exact manner in which you intend to use it. Most of us are rarely this lucky!

Search the literature, product reviews and product data sheets to determine how the antibody has been used in the past. Be mindful that antibodies that work well in one application may not be suited for another. For example, an antibody that works well in a Western may not work well in immunoprecipitations due to conformational changes in the protein. Alternatively, posttranslational modifications, such as phosphorylation, might mask or reveal epitopes recognized by antibodies (particularly monoclonal antibodies).

If you can’t find an antibody that has been used in your application, choose one that has been used in a compatible manner. For example, antibodies used in FACS analysis on live cells might also work in native immunoprecipitations which also maintain the integrity of the protein.

Check how the Amcyan antibody was validated

A lot of antibodies are validated using recombinant proteins that are abundantly expressed. Or they are validated with proteins that were produced in non-native hosts such as bacteria or yeast. While these validations show that the antibody can work, they do not necessarily show that the antibody will work under alternative conditions.

[caption id="attachment_18" align="aligncenter" width="120"] Western blot analysis of AmCyan fusion protein with anti-AmCyan tag monoclonal antibody(8T2) at 1:5000 (lane A) dilution.[/caption]

The type of antibody matters

Is it better to us a monoclonal or a polyclonal Amcyan antibody? For some proteins, you don’t have a say in the matter – you have to go with whatever antibody is available. However, if you do have a choice, choosing the right kind of antibody might increase your success. Polyclonal antibodies, which recognize multiple epitopes, are often better in Western blots, immunohistochemistry and immunoprecipitations. They can also be more robust and detect less abundant proteins.

Monoclonals are useful in detecting conformation-specific epitopes and are invaluable when using multiple antibodies simultaneously. Due to their specificity, monoclonals often give less background. Because they only recognize a single epitope though, monoclonal antibodies can be more limited in application. But don’t let that scare you off! I have used many monoclonal anbtibodies that worked well in Westerns.

If you are looking for a detection tool to quantify or localize your Amcyan epitope or Amcyan fusion proteins, we highly recommend you to try our featured monoclonal Amcyan Antibody risk free. This antibody with clone number 8T2 can specifically recognizes native and denatured forms of Amcyan fusion proteins, with which can meet your any requirements for Western Blot analysis.

Introduction to VSV-G antibodies from Sigma

One of the most frustrating things in the lab is spending good money on an antibody only to find out it doesn’t work well in your application. Now let start our VSV-G Monoclonal Antibody introduction from Sigma.

 Sigma-Aldrich Corporation, started from 1975, is an American chemical, life science and biotechnology company headquartered in St. Louis and has operations in approximately 40 countries. In September 2014, the German company Merck KGaA announced that it would be acquiring Sigma-Aldrich to found a new Millipore-Sigma. Due to this combination, Merck-Millipore would supply these antibodies from 2016 in your region.

Sigma (Now part of Millipore-Sigma) provide totally 2 VSV-G antibody, with one VSV-G polyclonal antibody and another VSV-G monoclonal antibody with clone #P5D4.

VSV-G antibody	Clonality	Host	Applications
ANTI-VSV-G TAG antibody produced in rabbit	Polyclonal	Rabbit	WB
Monoclonal Anti-VSV Glycoprotein antibody produced in mouse, clone P5D4	Monoclonal	Mouse	WB, IF, IP

The polyclonal VSV-G antibody can only be applied to WB assay with suggested 1:1000 dilution. So if you need VSV-G fusion protein localization analysis, you'd better choose another Monoclonal Anti-VSV Glycoprotein antibody. This monoclonal antibody recognizes an epitope containing the five carboxy-terminal amino acids of VSV Glycoprotein. In infected cells, the antibody localizes immature forms of VSV-G in the rough endoplasmic reticulum (RER) and in the cisternae of Golgi complex, as well as mature VSV-G at the cell epipolic and in the budding virus. It also can be used to pull down the VSV-G taged protein with IP assay.

As you know, one manufacturer may license their antibody to other companies, so do not buy three antibodies to test without making sure they are actually different. With common monoclonals that are supplied by many companies you might also consider shopping around for the best supplier of the target protein. Here, we put another high quality VSV-G Tag Mab from other vendor with Sigma VSV-G antibody together for your brief comparison.

Vendor	Clone #	Applications	Price Yr 2017
	P5D4	WB, IF, IP	$340/100μl $440/200μl
	14D2	WB, IF, IP	$90/50μl $260/200μl

This Anti-VSV-G-Tag Mouse Monoclonal Antibody (14D2) from Abbkine was affinity-purified from mouse ascites by affinity-chromatography using synthetic peptide immunogen containing the 15 carboxy-terminal amino acids (497-511) of VSV-G conjugated to KLH. So now you have two choice to get your VSV-G monoclonal antibody. If you're willing to saving cost or only need small size, Abbkine would be yours. Or, get this Sigma VSV-G Antibody from EMD-Millipore now with good brand awareness.

HA Tag DNA and Peptide Sequence

It is common to include a small segment of the viral hemagglutinin (HA) coat protein  in gene expression vectors as an epitope tag. Human influenza hemagglutinin (HA) is a surface glycoprotein required for the infectivity of the human virus. The HA tag has been extensively used as a general epitope tag in expression vectors. Many recombinant proteins have been engineered to express the HA tag, which does not appear to interfere with the bioactivity or the biodistribution of the recombinant protein. This tag facilitates the detection, isolation, and purification of the protein of interest.

As show below, Influenza Hemagglutinin (HA) Peptide is a tag peptide derived from an epitope of the influenza hemagglutinin protein. In conjugation with an HA hemagglutinin tag antibody, proteins tagged with the HA segment can be easily identified in Western blot or in immunocytochemical experiments, eliminating the generation of any protein-specific antibodies. Thus, HA hemagglutinin tag antibody is one of most commonly used antibodies in protein expression and cell biology studies.



 

However, the HA tag is not suitable for detection or purification of proteins from apoptotic cells since it is cleaved by Caspase-3 and / or Caspase-7 after its sequence DVPD, causing it to lose its immunoreactivity.

HA Tag DNA Sequence

Tags and fusion proteins are excellent tools for further understanding the function of your favorite gene. For example, fusing your protein to an epitope tag, such as HA, will allow you to easily identify your protein using an HA tag antibody against that epitope. This could allow you to conduct WB or IP of your favorite gene even if you do not have an antibody against it.

HA Epitope Vector	Representative Empty Backbones
pKH3 Vector	N or C-terminal 3xHA tag for mammalian expression
pHAHA Vector	N-terminal double HA tag for mammalian expression
pCS2HA Vector	N or C-terminal 2xHA tag in pCS for a variety of systems, including Xenopus
pAG423GPD-ccdB-HA	C-terminal 3xHA tag for yeast expression

DNA sequence coding for HA tag in pCMV-HA vector is from 832-858 with total 27 nucleotides, TAC CCA TAC GAT GTT CCA GAT TAC GCT 3' or 5' TAT CCA TAT GAT GTT CCA GAT TAT GCT.

VSV-G Tag Amino Acid Sequence

The HA tag is derived from the HA-molecule corresponding to amino acids 98-106, which is a 9 amino acid sequence YPYDVPDYA peptide derived from the Human influenza hemagglutinin.

Sequence	Tyr-Pro-Tyr-Asp-Val-Pro-Asp-Tyr-Ala
Chemical Name	Influenza Hemagglutinin Peptide
Canonical SMILES	CC(C)C(C(=O)N1CCCC1C(=O)NC(CC(=O)O)C(=O) NC(CC2=CC=C(C=C2)O)C(=O)NC(C)C(=O)O)NC( =O)C(CC(=O)O)NC(=O)C(CC3=CC=C(C=C3)O)NC (=O)C4CCCN4C(=O)C(CC5=CC=C(C=C5)O)N
Formula	C53H67N9O17	M.Wt	1102

If you are looking for a detection tool to quantify or localize your HA fusion proteins, we highly recommend you to try our featured monoclonal HA Antibody risk free. This antibody with clone number 4F6 can specifically recognizes native and denatured forms of HA fusion proteins, with which can meet your any requirements for Western Blot, Immunofluorescence and Immunoprecipitation assays.

Anti-CBP Tag Monoclonal Antibody(12H5) Review

CBP affinity tag is a useful tag in protein study which can be added to the N or C terminus of proteins of interest through DNA recombinant technology. The tag is derived from muscle myosin light-chain kinase. The tag comprises 26 amino acid residues with the molecular weight of 4 kDa and sequence of KRRWKKNFIAVSAANRFKKISSSGAL. CBP tag has the relatively high affinity for calmodulin (CaM). CBP-tagged proteins can be purified from crude cell extracts through CaM affinity resin. CBP Tag antibody is a useful tool in analysis of CBP-tagged proteins.

Anti-CBP Tag Monoclonal Antibody(12H5) was affinity-purified from mouse ascites by affinity-chromatography using specific immunogen. This antibody has been tested with WB. And Abbkine suggested starting dilutions are as follows: WB 1:5000.

The CBP tag binds to calmodulin resin in the presence of low concentration of calcium and can be eluted with a neutral buffer. The antibody is suitable for detecting CBP fusion proteins. The quality of the Anti-CBP Tag Monoclonal Antibody(12H5) is found satisfactory. I am very happy that I chose this product.

VSV-G Tag Sequence

VSV-G, a vesicular stomatitis virus G (VSV-G) protein fragment, which is commonly used in biomedical research to pseudotype retroviral and lentiviral vectors, conveying the ability to transduce a broad stroll of mammalian cell sign with genes of interest.

As show below, recognized VSV-G epitope represents the amino acid sequence derived from the Vesicular Stomatitis viral glycoprotein.



 

The VSV-G epitope tag is commonly engineered onto the N- or C- terminus of a protein of interest so that the tagged protein can be analyzed and visualized using immunochemical methods.

VSV-G Tag DNA Sequence

We cited VSV-G sequence information from Plasmid pCMV-VSV-G, which is from Dr. Bob Weinberg's lab published in RNA 2003 Apr;9(4):493-501. In this vector, VSV-G tag sequence coding envelope protein for producing lentiviral and MuLV retroviral particles has been used in conjunction with a packaging vector such as pCMV-dR8.2 dvpr (lentiviral) or pUMVC (MuLV retroviral).



DNA sequence coding for VSVG tag in pCMV-VSV-G vector is from 2936-2968 with total 33 nucleotides, TAT ACA GAC ATA GAG ATG AAC CGA CTT GGA AAG.

VSV-G Tag Amino Acid Sequence

VSV-G epitope tag is corresponding aa 501-511 of vesicular stomatitis virus glycoprotein (VSV-G), which is a 11 amino acid sequence YTDIEMNRLGK peptide derived from the Vesicular Stomatitis viral glycoprotein.

Sequence	Tyr-Thr-Asp-Ile-Glu-Met-Asn-Arg-Leu-Gly-Lys
Chemical Name	VSV-G Peptide
Canonical SMILES	CCC(C)C(C(=O)NC(CCC(=O)O)C(=O)NC(CCSC)C (=O)NC(CC(=O)N)C(=O)NC(CCCN=C(N)N)C(=O) NC(CC(C)C)C(=O)NCC(=O)NC(CCCCN)C(=O)O)N C(=O)C(CC(=O)O)NC(=O)C(C(C)O)NC(=O)C(CC1 =CC=C(C=C1)O)N
Formula	C57H94N16O19S	M.Wt	1334

If you are looking for a detection tool to quantify or localize your VSV-G fusion proteins, we highly recommend you to try our featured monoclonal VSV-G Antibody risk free. This antibody with clone number 14D2 can specifically recognizes native and denatured forms of VSV-G fusion proteins, with which can meet your any requirements for Western Blot, Immunofluorescence and Immunoprecipitation assays.

Multi-purpose PurKine™ Protein A/G Resin 4FF

Protein A/G is a genetically-engineered protein which combines the IgG binding domains of both Protein A and Protein G. The fusion protein is expressed in E. coli. Protein A/G contains four Fc-binding domains from Protein A and two from Protein G, making it a more general and convenient tool for investigating and purifying immunoglobulins. The surface is chemically modified with a proprietary method to minimize nonspecific binding of proteins. Also, Protein A/G binding to immunoglobulins is not as pH-dependant as Protein A.

PurKine™ Protein A/G Resin 4FF  is effective for affinity purification of IgG from serum, ascites fluid, cell culture supernantant and other antibody samples. It can also apply to immunoprecipitation (IP) and co-immunoprecipitation (Co-IP). The Resin consists of 90μm beads of cross-linked 4% agarose, to which Recombinant protein A/G has been coupled. Each  milliliter of drained resin will bind approximately 5-10 mg Human IgG. This product shows long term stability within pH range of 3 to 10.

PurKine™ Protein A/G Resin 4FF  binds to all human IgG subclasses and binds well to all mouse IgG subclasses but does not bind mouse IgA, IgM or serum albumin, making it ideal for purification of polyclonal or monoclonal IgG. In addition, the resin is very robust, which can tolerate up to 300cm/hour's  flow rates. No matter scientific research or industrial production, this resin will meet requirements.

Premium-quality PurKine™ Protein G Resin for antibody purification

Protein G, a bacterial cell wall protein isolated from group G Streptococci, binds to mammalian IgGs mainly through Fc regions. Native protein G has 3 IgG binding domains and also sites for albumin and cell-surface. Although protein G has very similar tertiary structures to protein A, their amino acid compositions differ significantly, resulting in different binding characteristics. Protein G can be used for purification of mammalian monoclonal and polyclonal IgGs that do not bind well to protein A.

PurKine™ Protein G Resin consists of 90μm beads of cross-linked 4% agarose, to which Recombinant protein G has been coupled. Recombinant Protein G, in which albumin and cell surface binding sites have been eliminated to reduce nonspecific binding, is produce in  E. coli. This affinity resin is of  high yield and high purity purification of whole IgG from mammalian serum and other fluids. Tests confirm that performance equals or exceeds popular recombinant protein G resins from other suppliers, and no decrease in performance occurs after at least five repeated uses.

PurKine™ Protein G Purification system has greater affinity than protein A for most mammalian IgGs, especially for certain subclasses including human IgG3,mouse IgG1 and rat IgG2a. Unlike protein A, protein G does not bind to human IgM, IgD and IgA. Per mL of the resin can capture more than 30mg human lgG. The resin is also available as prepacked column (PurKine™ Protein G Packed Column) and kit (PurKine™ Antibody Purification Protein G Kit) formats.

Anti-VSV-G-Tag Mouse Monoclonal Antibody (14D2) Review

VSV-G , a vesicular stomatitis virus G (VSV-G) protein fragment. VSV-G protein is commonly used in biomedical research to pseudotype retroviral and lentiviral vectors, conveying the ability to transduce a broad range of mammalian cell types with genes of interest. The VSIV G protein has also been used in cytological studies of trafficking in the endomembrane system. Immunoelectron microscopy suggests that VSIV G protein moves from cis to trans Golgi bodies without being transported between them in vesicles, supporting the cisternal maturation model of Golgi trafficking.

Anti-VSV-G-Tag Mouse Monoclonal Antibody (14D2) was affinity-purified from mouse ascites by affinity-chromatography using specific immunogen. This antibody has been tested with IF, IP and WB. And Abbkine suggested starting dilutions are as follows: WB 1:5000, IP 1:200, IF 1:1000.

Anti-VSV-G-Tag Mouse Monoclonal Antibody (14D2) has a high reputation with its wide application range and high reliability. We always have faith in the quality of Abbkine's products. As a useful and efficient product, Anti-VSV-G-Tag Mouse Monoclonal Antibody (14D2) is well worth to recommending.

PurKine™ Protein A Resin 4FF release

Protein A, a bacterial cell wall protein isolated from Staphylococcus aureus, binds to the Fc region of immunoglobulins through interactions with the heavy chain. For antibody affinity purification, recombinant protein A is a better choice. Recombinant protein A is produced in E. coli that contains five high affinity IgG binding domains with other non-essential domains removed to reduce nonspecific binding.

Abbkine PurKine™ Protein A Resin 4FF has been immobilized for use in IgG purification applications at a variety of scales. The Resin consists of 90μm beads of highly cross-linked 4% agarose, to which Recombinant protein A has been coupled. Per mL of resin can capture at least 40mg human lgG. In addition, its high flow properties (up to 300cm/hour) make it excellent for scaling-up.

PurKine™ Protein A Purification system is designed for easy, one-step purification of antibody from ascites fluid, serum (polyclonal antibody), or from the culture supernatant of a hybridoma cell line (monoclonal antibody). Protein A Resin 4FF is more suitable for large-scale industry purification to save time as well as energy. It can combine with  Peristaltic pump or AKTA instrument .

Anti-V5 Tag Mouse Monoclonal Antibody (11D5) Review

The recognized V5 epitope represents 95GKPIPNPLLGLDST108 of RNA polymerase α subunit of simian parainfluenza virus type 5. This short peptide sequence was chosen because high-affinity antibodies can be reliably produced in many different species. A 14-amino-acid V5 epitope derived from simian parainfluenza virus type 5 (Southernet al., 1991) was inserted into the C terminus of the capsid protein (before the stop codon ofORF2) to construct a recombinant marker virus. We demonstrated that the V5 epitope was displayed on the surface ofthe capsid protein. Furthermore, the recombinant marker virus behaved similarly to the parental virus in vitro and in mice, and could be differentiated from the parental virus via polymerase chain reaction (PCR) and serological methods.

Anti-V5 Tag Mouse Monoclonal Antibody (11D5) was affinity-purified from mouse ascites by affinity-chromatography using specific immunogen. This antibody has been tested with IF, IPand WB. And Abbkine suggested starting dilutions are as follows: WB 1:5000, IP 1:200, IF 1:1000.

Anti-V5 Tag Mouse Monoclonal Antibody (11D5) can be helpful in detecting the recombinant proteins, some of which include transmembrane and secreted proteins fusion protein. It applies to IF, IP and WB experiments. It's convenient to choose this product.

More cost-effective PurKine™ protein A resin

Protein A, a bacterial cell wall protein isolated from Staphylococcus aureus, binds to the Fc region of immunoglobulins through interactions with the heavy chain. For antibody affinity purification, recombinant protein A is a better choice. Recombinant protein A is produced in E. coli that contains five high affinity IgG binding domains with other non-essential domains removed to reduce nonspecific binding.

Abbkine PurKine™ Protein A Resin has been immobilized for use in IgG purification applications. The Resin consists of 90μm beads of cross-linked 4% agarose, to which Recombinant protein A has been coupled. Per mL of resin can capture more than 40mg human lgG. Tests confirm that performance equals or exceeds popular recombinant protein A resins from other suppliers. Column (PurKine™ Protein A Packed Column) which is prepacked with recombinant protein A resin is also supplied.

PurKine™ Protein A Purification system is designed for easy, one-step purification of classes, subclasses and fragments of immunoglobulins from biological fluids and from cell culture media. Protein A is generally preferred for affinity purification of rabbit, Guinea pig, dog and cat IgG. The portfolio allow optimization of process for maximum protein yield, stability and solubility. Tests also confirm that no decrease in performance occurs after at least five repeated uses.

Anti-RFP Tag Mouse Monoclonal Antibody (9D1) Review

RFP (Red Fluorescent Protein), cloned from Discosoma coral (DsRed, DsRed2 or drFP583), is a versatile biological marker for monitoring physiological processes, visualizing protein localization, and detecting transgenic expression in vivo. RFP can be excited by the 488 nm or 532 nm laser line and is optimally detected at 588 nm. It is common research practice for biologists to introduce a gene (or a gene chimera) encoding an engineered fluorescent protein into living cells and subsequently visualize the location and dynamics of the gene product using fluorescence microscopy.

Anti-RFP Tag Mouse Monoclonal Antibody (9D1) was affinity-purified from mouse ascites by affinity-chromatography using specific immunogen. This antibody has been routinely tested with WB application. And Abbkine suggested starting dilution is as follows: WB 1:5000.

I must thanks for the help of your product specialists. I was impressed by their professionalism and professional level. It is a wise choice to choose Abbkine.

 

Unsurpassed PurKine™ Strep II-Tag Streptactin Resin 4FF

The Strep-tag II is an eight-amino-acid peptide (WRHPQFGG), which, after fusion with the recombinant protein of interest, offers a practically useful solution to purify target protein. The Strep-tag II has negligible effect on the recombinant protein due to its chemically balanced amino acid composition. Generally, it does not interfere with folding or bioactivity and does not induce protein aggregation. Thus, there is no need for removing the tag.

PurKine™ Strep II-Tag Streptactin Resin 4FF effectively purifies high levels of Strep II-tagged proteins expressed in any express system such as, E. coli, insect, and mammalian cells at a variety of scales. The Resin consists of 90μm beads of cross-linked 4% agarose, to which Strep-Tactin has been coupled. Strep-Tactin is a streptavidin derivative which is one of the most stable proteins known. Tests confirm that performance equals or exceeds popular Strep-Tactin resins from other suppliers, and no decrease in performance occurs after at least five repeated uses. In addition, its high flow properties make it excellent for scaling-up.

PurKine™ Strep II-Tag Strep-Tactin Resin 4FF system is based on innovative high-capacity matrix for convenient single-step purifications of Strep II-Tag protein under physiological conditions, which means no influence of protein folding or function. The binding affinity of Strep II-Tag to Strep-Tactin is nearly 100 times higher than to streptavidin. Furthermore, the neutral pI of Strep-Tactin minimizes non-specific protein or nucleic acid binding. The product is safe and easy to use.

Anti-KT3 Tag Mouse Monoclonal Antibody (14D8) Review

The KT3 epitope tag is commonly engineered onto the N- or C- terminus of a protein of interest so that the tagged protein can be analyzed and visualized using immunochemical methods. KT3 epitope tag can be used to monitor expression of the protein products transfected with cDNA constructs. The recognized KT3 epitope represents the amino acid sequence KPPTPPPEPET derived from the Simian Virus 40 (SV40) large T-antigen.

Anti-KT3 Tag Mouse Monoclonal Antibody (14D8) was affinity-purified from mouse ascites by affinity-chromatography using specific immunogen. This antibody has been routinely tested with WB application. And Abbkine suggested starting dilution is as follows: WB 1:5000.

We always have faith in the quality of Abbkine's products. I chose Anti-KT3 Tag Mouse Monoclonal Antibody (14D8) in my test. I was impressed by the results. It's wonderful!

PurKine™ Biotin-Tag Streptavidin Resin 6FF Released !

Streptavidin is a 60kDa protein from Streptomycetes avidinii. The protein is a tetramer having four biotin-binding sites. Unlike avidin, streptvidin has a low isoelectric point (pI=5) and no carbohydrate groups, resulting in low nonspecific binding.

PurKine™ Biotin-Tag Streptavidin Resin 6FF is based on innovative high-capacity matrix for convenient single-step purifications of biotin and biotinylated substances. Biotinylated antibodies, proteins, peptides, nucleic acids and other molecules or interaction complexes can be captured, immunoprecipitated, removed or purified from samples using this streptavidin resin. The Resin consists of 90μm beads of cross-linked 6% agarose, to which Streptavidin has been coupled. Tests confirm that performance equals or exceeds popular Streptavidin resins from other suppliers, and no decrease in performance occurs after at least five repeated uses.

Per mL of streptavidin resin 6FF can capture 1 to 3 mg of biotinylated BSA or more than 120 nmol Biotin. Users can pack the resin themselves or they can purchase PurKine™ Biotin-Tag Streptavidin Packed Column 6FF which is pre-packed for convenient. In addition, streptavidin resin 6FF  is a highly crosslinked resin that is able to tolerate at most 500 cm/hour  linear flow rates. It's totally excellent for scaling up.

Affinity of Protein A, Protein G and Protein A/G

Protein A, Protein G and Protein A/G are often used in antibody purification and other applications such as immunoprecipitation (IP) and chromatin immunoprecipitation(ChIP). Their binding abilities to antibodies of different subtypes and species are varied. How to choose correct types of protein to purify your sample? Here, we list the antibody binding properties of Protein A, Protein G and Protein A/G.

Note: +++ strong binding; ++ medium binding; + weak binding; - no binding

Species	Antibody subtype	Protein A	Protein G	Protein A/G
Human	Total IgG	+++	+++	+++
	IgG1	+++	+++	+++
	IgG2	+++	+++	+++
	IgG3	+	+++	+++
	IgG4	+++	+++	+++
	IgM	+	-	+
	IgD	-	-	-
	IgA	+	-	+
Mouse	Total IgG	+++	+++	+++
	IgG2a	+++	+++	+++
	IgG2b	++	++	++
	IgG3	+	++	++
	IgM	-	-	-
Rat	Total IgG	+	++	++
	IgG1	+	++	++
	IgG2a	-	+++	+++
	IgG2b	-	+	+
	IgG2c	+++	+++	+++
Rabbit	Total IgG	+++	+++	+++
Goat	Total IgG	+	+++	+++
Sheep	Total IgG	+	+++	+++
Guinea pig	Total IgG	+++	+	+++
Hamster	Total IgG	++	++	++
Donkey	Total IgG	++	+++	+++
Horse	Total IgG	+	+++	+++
Cow	Total IgG	+	+++	+++
Pig	Total IgG	+++	+	+++
Dog	Total IgG	+++	+	+++
Cat	Total IgG	+++	+	+++
Chicken	Total IgY	-	-	-

Anti-mCherry Tag Mouse Monoclonal Antibody (9D3) Review

mCherry is a fluorophore (a fluorescent protein) used in biotechnology as a tracer to follow the flow of fluids, as a marker when tagged to molecules and cell components. mCherry and the majority of red fluorescent proteins derive from a protein isolated from Discosoma sp., while other fluorescent proteins in the green range are often variants of GFP from Aequorea victoria. mCherry is sometimes preferred to other fluorophores due to its colour, as well as its photostability compared to other monomeric fluorophores.

Anti-mCherry Tag Mouse Monoclonal Antibody (9D3) was affinity-purified from mouse ascites by affinity-chromatography using specific immunogen. This antibody has been routinely tested with WB application. And Abbkine suggested starting dilution is as follows: WB 1:5000.

The mCherry Tag Antibody can identify recombinant protein marked by mCherry that have alread been expressed, including those in the amino acid and the carboxyl terminal. Anti-mCherry Tag Mouse Monoclonal Antibody (9D3) worked very well. I am satisfied with the results of the experiment.

Easy use PurKine™ Biotin-Tag Protein Purification Kit (Streptavidin)

Streptavidin is a 60kDa protein from Streptomycetes avidinii. The protein is a tetramer having four biotin-binding sites. Unlike avidin, streptvidin has a low isoelectric point (pI=5) and no carbohydrate groups, resulting in low nonspecific binding.

PurKine™ Biotin-Tag Protein Purification Kit (Streptavidin) is based on innovative high-capacity matrix for convenient single-step purifications of biotin and biotinylated substances. Biotinylated antibodies, proteins, peptides, nucleic acids and other molecules or interaction complexes can be captured, immunoprecipitated, removed or purified from samples using this streptavidin resin. The Resin consists of 90μm beads of cross-linked 6% agarose, to which Streptavidin has been coupled. Tests confirm that performance equals or exceeds popular Streptavidin resins from other suppliers, and no decrease in performance occurs after at least five repeated uses.

Per mL of streptavidin resin can capture 1 to 3 mg of biotinylated BSA or more than 120 nmol Biotin. PurKine™ Biotin-Tag Protein Purification Kit (Streptavidin) contains PurKine™ Biotin-Tag Streptavidin Packed Column 6FF and buffers.There is no need for users to pack resin or prepare buffers themselves which can save precious time. In addition, because of the highly crosslinked resin, the tolerate linear flow rates of the kit is up to 500 cm/hour. It's totally ok for scaling up.

Anti-MBP Tag Mouse Monoclonal Antibody (9Y5) Review

MBP is used to increase the solubility of recombinant proteins expressed in E. coli. In these systems, the protein of interest is often expressed as a MBP-fusion protein, preventing aggregation of the protein of interest. The mechanism by which MBP increases solubility is not well understood. In addition, MBP can itself be used as an affinity tag for purification of recombinant proteins. The fusion protein binds to amylose columns while all other proteins flow through. The MBP-protein fusion can be purified by eluting the column with maltose. Once the fusion protein is obtained in purified form, the protein of interest (X) is often cleaved from MBP with a specific protease. Protein X can then be separated from MBP by affinity chromatography.The fusion of proteins with MBP usually enhances their solubility and facilitates their proper folding so that the fusion proteins are most often bifunctional. In addition, such fusions can facilitate the crystallisation of difficult proteins, e.g. membrane proteins.

Anti-MBP Tag Mouse Monoclonal Antibody (9Y5) was was affinity-purified from mouse ascites by affinity-chromatography using specific immunogen. This antibody has been tested with WB application. And Abbkine suggested starting dilutions are as follows: WB 1:5000.

Responsible to say, Anti-MBP Tag Mouse Monoclonal Antibody (9Y5) is a very good product. It works well. And this product is what I want.

PurKine™ MBP-Tag Dextrin Resin 6FF Products Released!

Dextrin Resins is a chromatography medium for purifying proteins fused to maltose binding protein (MBP-tagged protein). Recombinant proteins with MBP-tags often gives increased expression levels and higher solubility of the target protein. Proper folding of the attached protein has also been shown to be promoted by the MBP tag. Since MBP increase the solubility, the tag is extremely useful for recombinant proteins accumulated in an insoluble form (inclusion bodies).

PurKine™ MBP-Tag Dextrin Resin 6FF effectively purifies high levels of overexpressed MBP-tagged fusion proteins at a variety of scales. The Resin consists of 90μm beads of highly cross-linked 6% agarose, to which Dextrin has been coupled. Tests confirm that performance equals or exceeds popular Dextrin resins from other suppliers, and no decrease in performance occurs after at least five repeated uses. In addition, its high flow properties make it excellent for scaling-up.

The series of PurKine™ MBP-Tag Dextrin Resin products contain MBP-Tag Dextrin Resin 6FF, MBP-Tag Dextrin Packed Column 6FF, MBP-Tag Protein Purification Kit (Dextrin). Per milliliter of resin can capture more than 10 mg of MBP-tagged protein. MBP-Tag Dextrin Resin 6FF is a highly crosslinked resin which can tolerate at most 500cm/hour linear flow rates, thus it make  MBP-Tag Dextrin Resin 6FF more excellent for scaling-up.

Anti-Myc Tag Mouse Monoclonal Antibody (2D5) Review

A myc tag is a polypeptide protein tag derived from the c-myc gene product that can be added to a protein using recombinant DNA technology. It can be used for affinity chromatography, then used to separate recombinant, overexpressed protein from wild type protein expressed by the host organism. The peptide sequence of the myc-tag is (in 1- and 3-letter codes, respectively): N-EQKLISEEDL-C, N-Glu-Gln-Lys-Leu-Ile-Ser-Glu-Glu-Asp-Leu-C, where N stands for Amino-terminus and C stands for Carboxy terminus. It can be fused to the C-terminus and the N-terminus of a protein. The tag is approximately 1202 Daltons in atomic mass and has 10 amino acids.

Anti-Myc Tag Mouse Monoclonal Antibody (2D5) was affinity-purified from mouse ascites by affinity-chromatography using specific immunogen. This antibody has been tested with WB, IP and IF. And Abbkine suggested starting dilutions are as follows: WB 1:5000, IP: 1:200, IF: 1:1000.

Abbkine assures the top-ranking production technology and service system, and provides the quality products with logical prices. I trust the brand very much. Anti-Myc Tag Mouse Monoclonal Antibody (2D5) is one of the products that I think is very good.

New PurKine™ GST-Tag Glutathione Resin(Packed Column) 4FF

Glutathione S-transferase (GST) is a 26 kDa protein derived from Schistosoma japonicum. GST enzymes from various sources, both native and recombinantly expressed as fusion to the N-terminus of target proteins. Reduced glutathione (GSH), when immobilized as a ligand to agarose or other chromatography supports, enables high-yield and high-quality purification of recombinant proteins expressed as fusions with glutathione S-transferase (GST) or other glutathione binding proteins expressed in E. coli, insect cells and mammalian cells.

PurKine™ GST-Tag Glutathione Resin 4FF effectively purifies high levels of overexpressed GST-tagged fusion proteins at a variety of scales. The Resin consists of 90μm beads of cross-linked 4% agarose, to which reduced glutathione has been coupled. Tests confirm that performance equals or exceeds popular GST-Tag Purification Resins from other suppliers, and no decrease in performance occurs after at least five repeated uses. PurKine™ GST-Tag Glutathione Resin Packed Column 4FF is a column that is pre-packed with Glutathione Resin. There is no need to pack resin by users.

PurKine™ GST-Tag Purification system is based on innovative high-capacity matrix for convenient single-step purifications of recombinant glutathione S-transferase (GST) fusion proteins and other glutathione binding proteins. Per milliliter of resin can capture more than 10 mg of GST-tagged protein and target  proteins can be eluted under mild, non-denaturing conditions that preserve protein antigenicity and functionality. The difference between Glutathione Resin and  Glutathione Resin 4FF is the latter is a highly crosslinked resin which can tolerate at most 450cm/hour  linear flow rates, thus it make Glutathione Resin 4FF more excellent for scaling-up.

Anti-His Tag Mouse Monoclonal Antibody (5C3) Review

A polyhistidine-tag is an amino acid motif in proteins that consists of at least five histidine (His) residues, often at the N- or C-terminus of the protein. It is also known as hexa histidine-tag, 6xHis-tag, and by the trademarked name His-tag. Polyhistidine-tags are often used for affinity purification of polyhistidine-tagged recombinant proteins expressed in Escherichia coli and other prokaryotic expression systems.

Anti-His Tag Mouse Monoclonal Antibody (5C3)  was affinity-purified from mouse ascites by affinity-chromatography using specific immunogen. This antibody has been tested with IF, IP and WB. Abbkine suggested starting dilutions are as follows: WB 1:3000, IP: 1:200, IF: 1:1000.

His tag is a short peptide composed of 6 histidine (His-His-His-His-His-His). Because its molecular weight is small, His tag is relatively easy to isolate and purify. I chose Anti-His Tag Mouse Monoclonal Antibody (5C3) to detect the expression of His tag fusion protein. I am satisfied with the result.