Custom Polyclonal Antibody Production

Services / Antibody Services / Polyclonal Antibody

ABclonal has a comprehensive polyclonal antibody service that offers a personalized experience tailored to you. The antibodies produced will be validated by applications such as Western Blot and ELISA.

  • Polyclonal Antibody Production Process
Process Timeline
Antigen analysis and design If the customer provides the antigen: 10 weeks;
If ABclonal prepares the antigen:
14 weeks (protein antigen)
13 weeks (peptide antigen)
Antigen preparation Recombinant protein preparation
Peptide synthesis and conjugation
Potency test
Final bleed and serum purification
Validation WB
  • What We Need If:
ABclonal prepares 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).
You provide the 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.
ABclonal prepares a peptide antigen
Target gene sequence.
You provide the 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 there is an insufficient amount of antigen, Protein A or Protein G will be used to purify the antiserum.

The Host Species

ABclonal offers a variety of protocols (such as immunization schedules for rabbits, mice, rats, goats, etc.) for preparing custom polyclonal antibodies.

Our scientists will help you select the most suitable antibody service according to the host and your protein characteristic and research application.

Additional Services

One additional immunization and two blood collections for 80-100 mL of antisera.
Affinity purification of 25 mL of sera. Includes the affinity column.
On any bleed or purified antibody according to the customer's request.
Assay of antisera or purified antibodies according to the customer's request.
Assay of purified antibodies against tissue sections according to the customer's request.
Full range of custom peptide services available.
Full range of custom protein services available.

Professional Antigen Design (Free)MORE

Proper design of a suitable antigen is the most critical step in getting a useful antibody.

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.

High-quality antigen preparationMORE

According to your needs and the target characteristics, our experts will choose the best way to obtain the best immunogen.

Protein antigen

We experiment with a number of conditions (such as expression vector, host bacteria, and purification method) to improve the solubility, purity, and expression level. Through renaturation and refolding, we will ensure that the immunogen displays the correct epitopes, which improve the probability of the antibody recognizing the protein in its natural conformation.

Peptide antigen

For the most difficult-to-express antigens or those with low specificity, we choose the best region after a professional analysis, and synthesize a peptide (later coupled to KLH) as antigen to immunize the animals.

Our quality control procedures are very strict, ensuring that each synthetic peptide maintains high quality and high purity.

SPF animals

Specific-pathogen-free animals will be used for immunization.

Antigen affinity purification

Antibodies are purified by Protein A and antigen affinity chromatography.
When needed, ABclonal can provide cross-purification to obtain more specific antibodies.

  • Provides approximately 50-80 ml of antiserum per animal.
  • Produces antibodies with higher specificity

Antibody validation

Guaranteed results:

  • Affinity purified custom polyclonal antibody.
  • 1 ng of protein antigen or 100 ng of peptide antigen is detected when the antibody is diluted at 1:1000.

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 (5 mL per rabbit)
Primary Injection Day 1 Immunize with 0.50 mg of antigen in CFA, 10 SQ sites
1st Booster Day 14 Boost with 0.25 mg of antigen in IFA, 4 SQ sites
2nd Booster Day 28 Boost with 0.25 mg of antigen in IFA, 4 SQ sites
Serum Collection Day 35 Bleed (-25 mL per rabbit)
3rd 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
Note: After Day 35, the scheduled days in this protocol are approximations and are dependent on the titer test.

High-quality scientific research papers have been successfully published in well-known journals regarding ABclonal antibodies. We will continue to offer outstanding custom polyclonal antibody services as an effort to service the scientific research community at large.

  1. For quotation requests and questions, you may contact us by email, phone, or via our online quotation system.
  2. Orders can be placed by phone, email, fax, or online with a formal Purchase Order or credit card.
  3. Please send your samples to ABclonal, 86 Cummings Park Dr, Woburn, MA 01801, United States.