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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—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)
2-rat 80-day protocol

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 (± 2 days).

Procedure Protocol day Description
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

  • 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 (exsanguinatory bleed)
Protocol extension

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.

Procedure Protocol day Description
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

  • 1 vial: 1 mL test bleed post-inject sera
Resources

For Research Use Only. Not for use in diagnostic procedures.