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.
|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|
|Final bleed and serum purification|
If there is an insufficient amount of antigen, Protein A or Protein G will be used to purify the antiserum.
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.
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.
According to your needs and the target characteristics, our experts will choose the best way to obtain the best immunogen.
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.
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.
Specific-pathogen-free animals will be used for immunization.
Antibodies are purified by Protein A and antigen affinity chromatography.
When needed, ABclonal can provide cross-purification to obtain more specific antibodies.
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.
|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|
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.