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.

Highlights

  • Free Antigen Design

    • Based on experience, data, and structural characteristics of your desired target
    • Tailored to your specified experimental applications
    • Artificial intelligence (AI)-assisted algorithm to balance affinity and immunogenicity
  • >95% Success Rate

    >95% of projects produce antigen-specific antibodies.

    Highly Experienced

    >More than 100,000 polyclonal antibodies generated.

Publications

High-Impact scientific research papers have been successfully published in well-known journals regarding ABclonal antibodies, including Cell, Nature, and Science.

Polyclonal Antibody Production Process

Antigen Preparation

We immunize rabbits, mice, and rats 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.

  • If you decide to provide your own recombinant protein antigen

    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.

  • If you want us to prepare a recombinant protein antigen

    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).

  • If you decide to provide your own peptide antigen

    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.

  • If you want us to prepare a peptide antigen

    Antigen peptide sequence.

If there is an insufficient amount of antigen, Protein A or Protein G will be used to purify the antiserum.

Immunization Schedule

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:

1 WEEK

Primary Injection

  • Preimmune bleed
  • Initial immunization

7-8 WEEKS

Booster

  • Three booster doses

1-2 WEEKS

Test Bleed Collection

  • Post-booster antiserum testing by ELISA

1-2 WEEKS

Final Collection

  • Sacrifice rabbits for terminal bleed (40-50 mL/ rabbit)
  • Additional boosters available if desired

Note: The scheduled weeks in this protocol are approximations and are dependent on the titer test.

Polyclonal Antibody Purification

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.

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