Find out everything you need to know about our products here.
Our antibodies are guaranteed for 1 year after purchase. However, our internal studies show that the antibody will remain functional for a minimum of 5 years if properly stored.
Please store the antibody in its original tube at -20°C. Our antibodies are stored in 50% glycerol, which prevents them from freezing. Aliquoting is unnecessary for -20°C storage.
Yes! Feel free to email us with the product number in the subject line. Alternatively, when you are on a specific product page, you can find a printable datasheet by clicking the Print Icon below the product number at the top of the page (Figure 1).
The concentration of a given antibody is found on the product datasheet and on the product vial tube itself. In the event that the concentration displayed on the vial tube is different to that shown on the product datasheet, please refer to the vial tube concentration.
On the antibody product page, please click the link beside ‘Immunogen’ under the ‘Immunogen Information’ section (Figure 2) to view the fusion protein with the amino acid sequence (Figure 3). Should you require more detailed information, please speak to a member of our staff via live chat.
Yes, we offer blocking proteins for your experiments. A link to these products can be found beside ‘Immunogen’ under the ‘Immunogen Information’ section of the antibody product page.
We usually test our antibodies on human and mouse cells or tissues. However, our guarantee covers the antibody in any species.
All currently tested applications are listed on each product page. Should you require more details or information regarding an application other than those listed, please contact our team.
This depends on the extent of protein sequence similarity between the immunogen and the potential cross-reactive protein sequence. A pair-wise sequence alignment can be performed online through the NCBI-BLAST website.
Each product page shows the number of publications and has links to the publication if the antibody has already been cited (Figure 4).
The choice of appropriate secondary antibody is determined by the specifications of the primary antibody and the application (i.e., species, subclass, and fragment type).
Guiding principles for choosing a suitable secondary antibody:
Proteintech offers a variety of secondary antibodies suitable for Western Blotting, ELISA, cellular imaging, and flow cytometry. View our secondary antibodies catalog for more detailed information.
Isotype controls are used to determine non-specific interactions in a given sample such as Fc receptor binding. Most isotype controls are monoclonal and most are not suitable for use with polyclonal antibodies as they contain more than one IgG class. The isotype control antibody should match the primary antibody regarding host species, isotype, and conjugation. Lastly, the sample incubated with the isotype control antibody and the sample incubated with the primary antibody should always be run in parallel.
A sample without a primary antibody can be used as a negative control to distinguish specific and non-specific signal from the secondary antibody. Any products/stainings obtained with these control conditions can be attributed to non-specific (off-target) interactions. Furthermore, samples that have been genetically manipulated to alter the target’s levels (e.g., CRISPR) can be used as a negative control.
For a positive control, any tissues, cells, or lysates that have been used successfully in our validation data set or by our customers are a suitable control. Each antibody has its own PubMed publication record on our website. Additionally, the Uniprot, Omnigene, and GeneCards databases are great resources for finding tissues/cell lines with high expression of the target. The latest publications record in PubMed can also be a useful tool for detailed and the most up-to-date information regarding target research.
Each monoclonal antibody has a clone number that is specific for its single clone of hybridoma cells. The clone number can be found on the product page next to the catalog number (Figure 5).
There may be several reasons for a difference in band size, such as multimers, gel migration, relative charges, splice variants, post-translational modification, or cleavage. For more information, please contact our technical support.
Yes, it is recommended to perform an antigen retrieval step since the fixation process during paraffinization cross-links proteins and may mask the epitopes, resulting in weak or false negative staining. However, this situation can be rectified with a heat-induced epitope retrieval (HIER) or proteolytic-induced epitope retrieval (PIER) step. The most suitable choice depends on the tissue type and primary antibody.
We usually add 6x His tags or GST tags to the N-terminus of proteins; these proteins are then purified by Ni2+ and glutathione affinity chromatography, respectively.
The His-tagged recombinant protein is usually stored in 1x PBS buffer (58mM Na2HPO4, 17mM NaH2PO4, 68mM NaCl, pH 7.4) containing 300mM imidazole.
The GST-tagged recombinant protein is usually stored in 1x PBST buffer (58mM Na2HPO4, 17mM NaH2PO4, 68mM NaCl, 1% Triton-X100, pH 7.4) containing 100mM glutathione (GSH).
Antibody-coated 96-well microplate
Tetramethylbenzidine (TMB) substrate
Plate cover seals
All Proteintech ELISA kits are two-site sandwich ELISAs.
Yes, all plates are pre-coated with an antibody that is specific for the target.
It is recommended to run the experiment in duplicate at a minimum. Triplicate would be ideal.
No, different lot numbers cannot be mixed.
Incubation times should be followed exactly as stated in the manual.
Proteintech ELISA kits can be stored at 4C for 6 months after receipt.
Most of our ELISA kits are validated for cell culture supernatant, plasma, or serum. However, the Proteintech Guarantee covers all sample types.
The ELISA sensitivity is highlighted in the data sheet for each kit.
There are multiple explanations: incorrect storage, incorrect incubation time, improper antibody, insufficient washing, etc. For more information, please contact technical support.
Improper washing, contaminated reagents, samples, or wells. For more information, please contact technical support.
Custom production projects can typically take up to 6 months to complete, but due to the complexity of the process, we cannot always give a guarantee of an exact schedule. Please see below for the schedule of our 102-day immunization protocol, which demonstrates our commitment to making sure that you receive an antibody that works.
102-Day Immunization Protocol
|Day 0 Pre-Immune Bleed||Day 56 Test Bleed & ELISA||Day 88 Production Bleed|
|Day 1 Primary Injection||Day 60 Boost 3||Day 102 Final Bleed|
|Day 28 Boost 1||Day 74 Production Bleed|
|Day 42 Boost 2||Day 78 Boost 4|
No, unfortunately we do not currently accept fusion proteins expressed by our customers.
KLH is the default carrier protein. This is linked to the peptide via a terminal cysteine (added by Proteintech®). If you have cysteine residue in the middle of your peptide then we can use MAP beads instead.
Each antigen will be injected into two rabbits.
In our experience, full-length fusion protein antigens usually generate antibodies with higher titers and higher sensitivities. We recommend the full-length fusion protein procedure if your protein of interest has low sequence similarity with other proteins. However, if you need an antibody that is specific to a certain protein, the sequence of which is highly similar to other proteins, we strongly recommend using a peptide antigen. Other reasons you might choose a peptide antigen include:
1. Not having the cDNA for the protein of interest;
2. Expression of the full-length fusion protein has not been successful for some reason.
No, we carry out custom production of antibodies for proteins from any species, depending on antigenicity.
Each rabbit will yield approximately 12 ml of serum in each production bleed and 45–50 ml of serum from 90–100 ml of blood in the final bleed, totalling about 150 ml of antiserum from two rabbits.
We decide that based on the ELISA results with the test bleeds. If both rabbits give good results, we will purify 15 ml of serum from each rabbit. If one rabbit gives better results than the other, we will purify 30 ml of serum from this rabbit.
It depends on the concentration of the specific antibody in the serum provided. Usually we can get 2–10 ml of purified antibody from 15 ml of antiserum.
Please visit the custom production section for more information on the service we provide.