Intended use: standard generation of murine monoclonal and polyclonal antibodies (ELISA titers up to 1:10000~1:1000000) by two immunizations within 5 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 21 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 35 post-priming and measure the antibody titers by ELISA. ELISA titers should be in the range of 1:10000~1:1000000. After that the whole serum can be collected or proceed to antigen boost and cell fusion according to conventional hybridoma protocols.
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