How should antigen be prepared for immunisation?
Antigen should be supplied in buffered (Tris, MOPS, Hepes) saline solution. Insoluble antigen (inclusion bodies) is as good as a soluble one. Desired antigen concentration is 1 mg/ml, however lower concentrations are also acceptable. You can also send us just a band cut out from SDS-PAGE gel, which contains protein of interest. Destain gel in water, since acetic acid will contribute to further denaturation of the protein. • Avoid: Additives toxic to animals, e.g. protease inhibitors such as PMSF, sodium azide etc. • Acceptable: Low amounts of SDS, imidazole, urea, guanidine, non-ionic detergents, EDTA or EGTA and polyacrylamide. Final amounts depend very much from sample volume/antigen concentration. Please, inquire.
