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Custom Polyclonal Antibody Production

Free antigen design for customized application-specific polyclonal antibody.

We provide customized services for producing application-specific polyclonal antibodies used in your research. Leverage our years of experiences 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 characteristics of your desired target. learn more

Step 1: Cross-examine the sequence provided by the customer with the information in multiple databases, such as NCBI.

Step 2: Screen suitable, immunogenic protein or peptide candidates.

Step 3: Compare BLAST searches with genome or protein databases to identify ideal protein or peptide candidates.

Step 4: Perform protein alignments of homologs/orthologs/isoforms.

Step 5: Refer to relevant publications to better understand the 3D structure of the protein and the protein-protein/protein-DNA interaction.
Wider Choice of Host Animals – Choose from SPF (specific-pathogen-free) host animals, including rabbit, mouse, rat, and guinea pig.
High Specificity – Antibodies are purified by Protein A and antigen affinity chromatography. When needed, ABclonal can provide cross-purification to obtain a more specific antibody.
Guaranteed Result – 1 ng of protein antigen or 100 ng of peptide antigen is detected when the antibody is diluted at 1:1000.

Polyclonal Antibody Production Process

Antigen Design

We immunize rabbits, mice, rats, and guinea pigs to generate custom polyclonal antibodies against peptide antigens and protein antigens. Both peptide and protein antigens can be provided by you or prepared by us.

If you decide to provide your own recombinant protein antigen

More than 4 mg of protein antigen with purity above 80% and concentration above 1 mg/ml (immunization requires 3 mg and antigen affinity purification requires more than 4 mg of antigen); protein SDS-PAGE electrophoresis map.

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; constructed expression plasmid or expressed bacteriophage (expression conditions and protein SDS-PAGE electrophoresis map).

If you decide to provide your own peptide antigen

More than 10 mg of peptide antigen with purity above 85% (immunization requires 4 mg and antigen affinity purification requires more than 6 mg of antigen);HPLC and MS report.

If you want us to prepare a peptide antigen

Target gene 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 in the shortest possible time. It is the most affordable option and a popular choice among researchers.

The protocol uses two New Zealand White rabbits that are specific-pathogen-free. Injections are subcutaneous (SQ) as emulsions in Complete Freund's Adjuvant (CFA) or Incomplete Freund's Adjuvant (IFA). We guarantee that the same antigen will be injected into both animals.

Procedure	Protocol Day	Description
Control Serum Collection	Day 0	Pre-immune bleed (1 mL per rabbit)
Primary Injection	Day 1	Immunize with 0.50 mg of antigen in CFA, 10 SQ sites
1^st Booster	Day 14	Boost with 0.25 mg of antigen in IFA, 4 SQ sites
2^nd Booster	Day 28	Boost with 0.25 mg of antigen in IFA, 4 SQ sites
Serum Collection	Day 35	Bleed (-25 mL per rabbit)
3^rd Booster	Day 42	Boost with 0.25 mg of antigen in IFA, 4 SQ sites
Serum Collection	Day 56, 58	Two bleeds (total of -50 mL per rabbit)
ELISA Test	Day 60	ELISA titration (results available online) Verify disposition of rabbits Decide to continue or terminate
Purification	Optional	6-10 mg of purified antibodies