Tag and Loading Control Antibodies
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We provide customized services for producing application-specific polyclonal antibodies used in your research. Leveraging our years of experience in polyclonal antibody production: from antigen design, immunogen preparation, antibody development, antibody purification, modification, validation, to immunoassay development.
Based on experience, data, and structural characteristics of your desired target.
>95% of projects produce antigen-specific antibodies.
>More than 100,000 polyclonal antibodies generated.
High-Impact scientific research papers have been successfully published in well-known journals regarding ABclonal antibodies, including Cell, Nature, and Science.
We immunize rabbits, mice, rats, and guinea pigs to generate custom polyclonal antibodies against peptide and protein antigens. Both peptide and protein antigens can be provided by you or prepared by us. Below are the antigen requirements for standard rabbit polyclonal antibody development.
At least 4 mg of protein antigen with purity above 80% and concentration above 0.5 mg/ml (immunization requires 3 mg and antigen affinity purification requires additional 4 mg of antigen); protein SDS-PAGE image.
Target gene sequence and plasmid templates (more than 20 µl at 100 ng/µl concentration). We can also synthesize the gene if templates are not available; construct expression plasmid, pilot expression and scale up (expression conditions and protein SDS-PAGE image).
At least 5 mg of peptide antigen with purity above 85% (immunization requires 4 mg and antigen affinity purification requires additional 8 mg of antigen); HPLC and MS report.
Target gene sequence.
If there is an insufficient amount of antigen, Protein A or Protein G will be used to purify the antiserum.
ABclonal's standard protocol for polyclonal antibody is optimized to produce antiserum containing effective antibodies in the shortest possible time.
Our standard rabbit immunization protocol uses two New Zealand White rabbits that are specific-pathogen-free. Rabbits are immunized by subcutaneous injections of antigen mixed with Complete Freund's Adjuvant (CFA) or Incomplete Freund's Adjuvant (IFA). The standard protocol includes 4 injections for protein antigen and 5 injections for peptide antigen. Immunization schedule might be slightly different based on performance site.
Below is an example of using protein antigen for immunizing two rabbits:
|Control Serum Collection||Day 0||Pre-immune bleed (2 mL/rabbit)|
|Primary Injection||Day 1||Immunize with 400μg of antigen in CFA, SQ|
|1st Booster||Day 14||Boost with 200μg of antigen in IFA, SQ|
|2nd Booster||Day 35||Boost with 200μg of antigen in IFA, SQ|
|3rd Booster||Day 56||Boost with 200μg of antigen in IFA, SQ|
|Test Bleed Collection||Day 63||Post-4th immunization antiserum (2 mL/ rabbit) titer testing by ELISA against antigen|
|Terminal Bleed Collection/ Additional booster||Day 70||Sacrifice rabbits to collect terminal bleed (40-50 mL/ rabbit) or continue immunization with additional booster|
Note: After Day 63, the scheduled days in this protocol are approximations and are dependent on the titer test.
Depending on your requirement for specificity and yield, we offer two purification methods: Protein A purification and antigen affinity chromatography. In addition, we can provide cross-purification to obtain a more specific antibody upon request.