Standard 70-day rabbit immunization protocol for rabbit polyclonal antibody production
Abbkine has offered polyclonal antibody manufacturing services for more than four decades. Our expertise includes using a variety of different hosts for the development of polyclonal antibodies, including the most commonly used New Zealand white rabbits. Goats, sheep, rats, mouse and donkeys are also often selected.
This 2-Rabbit 70-Day protocol is optimized to produce antiserum in the shortest possible time. It is the most affordable option, popular among individual academic researchers.
The default immunization schedule uses two NZW SPF rabbits (New Zealand White rabbits that are Specific Pathogen-Free); additional rabbits are easily added to the standard order for a modest cost. Injections are subcutaneous (SQ) as emulsions in Complete Freund’s Adjuvant (CFA) or Incomplete Freund’s Adjuvant (IFA); alternative adjuvants can be used if requested. Continuation of rabbits after Day 60 is offered on a month-to-month basis. A brief protocol is listed below.
Antigen preparation, such as peptide synthesis and/or immunogen conjugation, occurs before Day 0. Injection amounts are given for a conjugated peptide antigen (e.g., KLH) or protein immunogen; MAP-peptide antigens are injected at 0.5mg throughout. Protocol Days are approximate (Â± 2 days).
|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 (~50 mL total per rabbit)|
|ELISA and shipping||Day 60||ELISA titration (results available online);
Verify disposition of rabbits; decide to continue or terminate
|Instruction due date||Day 72||If no animal instructions are received, per diem charges will apply|
|Deliverables (Day 60): 12 vials crude antibody sera