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Immunization and serum-collection procedures for custom rat polyclonal antibody production

Our comprehensive array of custom antibody services includes a customizable option for rat immunization, serum-collection, and titer analysis for rat antibody production, plus extension possibilities.

Custom Antibody Request

Highlights

  • Custom rat antibodies—a reliable protocol for immunizing rats, collecting serum, testing bleeds, reporting titer results, and delivering antisera
  • Full-service integration—combine this immunization service with our complete set of services for antigen preparation and antibody purification

This rat immunization schedule uses two Sprague Dawley rats for each 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 upon request.

80-Day 2 Rat Immunization Protocol

Antigen preparation, such as peptide synthesis and/or immunogen conjugation, occurs before Day 0. Protocol days are approximate (± 2 days).

ProcedureProtocol dayDescription
Control serum collectionDay 0Pre-immune bleed (~1 mL per rat)
Primary injectionDay 1Immunize with 100 µg antigen in CFA, SQ 4 sites
1st boosterDay 21Boost with 50 µg antigen in IFA, SQ 4 sites
2nd boosterDay 42Boost with 50 µg antigen in IFA, SQ 4 sites
Test bleedDay 50Test bleed (~1 mL per rat)
ELISA and shippingDay 60ELISA titration of pre-immune and test bleed;
(results available online)
3rd boosterDay 62Boost with 50 µg antigen in IFA, SQ 4 sites
DecisionDay 64Verify disposition of rats; decide to continue or terminate
Instruction due dateDay 80Terminal bleed (exsanguination, yielding 2–4 mL per rat)
and termination of animals if instructions not received

Deliverables: 6 vials crude antibody sera

  • 2 vials: 1 mL test aliquots pre-immune sera
  • 2 vials: 1 mL test bleeds post-inject sera (Day 50)
  • 2 vials: 2–4 mL terminal bleeds (exsanguination bleed)