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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 Antibody Request

Highlights

  • Custom rat antibodies—a reliable protocol for immunizing rats, collecting serum, testing bleeds, reporting titer results and delivering antisera
  • Track and customize project—track and manage projects online with the OpenProject Tool: you decide which animals to continue or terminate
  • 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 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 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).

Procedure Protocol day Description
Control serum collection Day 0 Pre-immune bleed (~1 mL per rat)
Primary injection Day 1 Immunize with 100 µg 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

  • 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)