Custom Rat Polyclonal Antibody Production Protocols
Immunization and serum-collection procedures for custom rat polyclonal antibody production
Our comprehensive array of Thermo Scientific Pierce custom antibody services includes a customizable option for rat immunization, serum-collection and titer analysis for rat antibody production, plus extension possibilities.
- Custom rat antibodies—a reliable protocol for immunizing rats, collecting serum, testing bleeds, reporting titer results and delivering antisera
- Track and customize project—easily add additional rats to the default protocol; track and manage projects online with OpenProject Tool: you decide which animals to continue or terminate (see Extensions below)
- Full service integration—combine this immunization service with our complete set of services for antigen preparation and antibody purification (see Home or Custom Polyclonal Antibodies pages for details)
This rat immunization schedule uses two Sprague Dawley rats for each MAP-peptide, conjugated-peptide antigen, or protein immunogen. Animals are housed individually in separate cages. 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.
80-day 2-rat immunization protocol for rat polyclonal antibody production
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), MAP-peptide, or protein immunogen. Protocol Days are approximate (Â± 2 days).
|Control serum collection||Day 0||Pre-immune bleed (~1 mL per rat)|
|Primary injection||Day 1||Immunize with 0.10 mg antigen in CFA, SQ 4 sites|
|1st booster||Day 21||Boost with 50 µg antigen in IFA, SQ 4 sites|
|2nd booster||Day 42||Boost with 50 µg antigen in IFA, SQ 4 sites|
|Test bleed||Day 50||Test bleed (~1 mL per rat)|
|ELISA and shipping||Day 60||ELISA titration of pre-immune and test bleed;
(results available online)
|3rd booster||Day 62||Boost with 50 µg antigen in IFA, SQ 4 sites|
|Decision||Day 64||Verify disposition of rats; decide to continue or terminate|
|Instruction due date||Day 80||Terminal bleed (exsanguination, yielding 2-4 mL per rat)
and termination of animals if instructions not received
Deliverables: 6 vials crude antibody sera
A pre-defined (yet customizable) option is available for extending or continuing the rat immunization and serum-collection schedule. The purchase decision to continue antibody production can be made at any time before scheduled termination. The terminal exsanguination bleed is included for no cost as part of the default protocol.
One-month extension protocol for rat antibody production.
|1st booster||Day 1||Boost with 0.10 mg antigen in IFA, 4 SQ sites|
|Serum collection||Day 14||Bleed (1-2 mL per rat)|
Deliverables (per rat): 1 vial crude antibody sera
Tools and information
For Research Use Only. Not for use in diagnostic procedures.