Immunization and serum-collection procedures for producing custom mouse polyclonal antibodies

Our array of Custom Antibody Services includes a customizable option for mouse immunization, serum-collection, and titer analysis for mouse polyclonal antibody production.

Custom Antibody Request

Highlights

  • Custom mouse antibodies—a reliable protocol for immunizing mice, 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

5-mouse 80-day immunization protocol

This mouse immunization schedule uses five BALB/c mice 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. Customizations to the protocol are available. For example, you can choose the number of mice needed and you can adjust the length of your protocol beyond the 80-day immunization schedule to meet your research goals. Please contact us to discuss the options in detail. Antigen preparation, such as peptide synthesis and/or immunogen conjugations, occur before day 0. Protocol days are approximate (+ 2 days).

ProcedureProtocol DayDescription
Control serum collectionDay 0Pre-immune bleed (0.2-0.5 mL per mouse)
Primary injectionDay 1Immunize with 0.10 mg 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 (0.2-0.5 mL per mouse)
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 mice; decide to continue or terminate
Instruction due dateDay 80Terminal bleed (exsanguination, yielding 0.5-1 mL per mouse)
and termination of animals if instructions not received

Deliverables: 15 vials crude antibody sera

  • 5 vials: 0.1 mL test aliquots pre-immune sera
  • 5 vials: 0.1 mL test bleeds post-inject sera (Day 50)
  • 5 vials: 0.5-1 mL terminal bleeds (exsanguinatory bleed)