3 mice/antibody x 100 µg antigen/mouse = 300 µg antigen/boost
4 -5 boosts x 300 µg antigen/boost = 1.2 – 1.5 mg antigen
This is the minimum amount of antigen needed for injection.
The cleaner the antigen, the better the response.
For mice, antigen concentrations of 2 mg/ml or higher are preferred because antigen gets diluted with Freund’s Adjuvant for the injection*. Maximum injection volume is ~150 µl, but we prefer injecting ~100 µl.
*NOTE: Antigen can also be left bound to beads. If you do not elute antigen,
it is necessary to wash the resin thoroughly with PBS. Antigen bound to
beads must also be as close to 2 mg/ml (or higher) as possible.
If we end up making monoclonal antibodies, we will need at least 1 mg antigen for ELISA screening.
Mouse injections are usually performed every 2 weeks with bleeds ~10 - 12 days post injection.
