ABclonal provides a reliable and cost-efficient custom polyclonal antibody service in SPF rabbit, mouse, rat, goat, chicken and guinea pig for gene-specific detection reagents. Our polyclonal antibodies can be used in multiple applications, including Western blots, ELISA, and IHC. However, developing highly specific and sensitive polyclonal antibody production is challenging and requires expertise during every step. Our team utilizes state-of-the-art scientific advances and the best tools to provide the highest quality polyclonal antibody service available.
1. If the antigen is provided by the customer: 5 mg of peptide or 5 mg of protein, purity >90% is required. The concentration should be >0.4 mg/ml, and the antigen solution must not contain organic solvents.
ABclonal offers a variety of protocols (immunization schedules for rabbits, mouse, rat, goats, etc.) for preparing custom polyclonal antibodies.
Our scientists will help you select the best antibody service according to the host and your protein characteristic and research application.
Proper design of a suitable antigen is the most critical step of getting an useful antibody.
According to the customers’ demand and the target characteristics, our experts choose the best way to get the best immunogen.
We experiment with a number of conditions to improve the protein expression level, solubility, and purity, such as expression vector, host bacteria, and various purification methods. Through renaturation and refolding, the immunogen displayed correct epitopes, which improved the probability of antibody recognition of 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.
Pure SPF (Specific pathogen Free) class of experimental animals are used for immunization.
According to the needs of customer and the titer test results of post-fourth immunization anti-serum, the immunization cycle is adjusted appropriately.
Antibodies are purified by protein A and antigen affinity chromatography.
Cross-purification can be provided by ABclonal to obtain more specific antibodies when needed.
ABclonal's standard option for polyclonal antibody is optimized to produce antiserum in the shortest possible time. It is the most affordable option, popular 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 a single antigen will be injected into both animals.
|Control Serum Collection||Day 0||Pre-immune bleed (5mL per rabbit)|
|Primary Injection||Day 1||Immunize with 0.50mg of antigen in CFA, 10 SQ sites|
|1st Booster||Day 14||Boost with 0.25mg of antigen in IFA, 4 SQ sites|
|2nd Booster||Day 28||Boost with 0.25mg of antigen in IFA, 4 SQ sites|
|Serum Collection||Day 35||Bleed (~25mL per rabbit)|
|3rd Booster||Day 42||Boost with 0.25mg of antigen in IFA, 4 SQ sites|
|Serum Collection||Day 56, 58||Two bleeds (~50mL total 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 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 with in an effort to service the scientific research community at large. Our goal is always to provide our customers with "the Better Antibody".