Intended use: rapid generation of murine monoclonal and polyclonal antibodies (ELISA titers up to 1:10000~1:100000 or higher) by two immunizations within 3 weeks.
1. Dilute the antigen with saline to 2-fold final concentration (50μl per injection).
Note: Recommended antigen dosage is: (1) 5~20μg per injection for low immunogenic antigens such as recombinant subunit proteins or 20~50μg for peptide-carrier conjugates; (2) 1~5μg per injection for high immunogenic antigens such as inactivated viruses or recombinant virus-like particles.
2. Mix the adjuvant by vortexing, sterilely remove the required volume (50μl per injection), and mix with the antigen at 1:1 ratio (v/v) as quick as possible.
Note: It is normal to see precipitation in the adjuvant.
3. Inject 100μl of the antigen/adjuvant mixture into a quadriceps muscle of each mouse.
Note: It is normal to see precipitation in the antigen/adjuvant mixture. Please mix well before drawing into syringe and inject as quick as possible after drawing into syringe.
4. Boost animals on day 14 post-priming.
Note: For boosting, please follow steps 1-3 of the priming protocol, using freshly prepared antigen/adjuvant mixture.
5. Bleed from tail tips on day 21 post-priming and measure the antibody titers by ELISA. ELISA titers should be in the range of 1:10000~1:100000. After that the whole serum can be collected or proceed to antigen boost and cell fusion according to conventional hybridoma protocols.
If you have any questions, please feel free to let us know: firstname.lastname@example.org, thanks.