How should the antigen be prepared for immunisation?
Proteins should be supplied in buffered saline solution (HEPES, phosphate, Tris-HCl, MOPS, etc.), as a lyophilizate or as a polyacrylamide gel slice. The slice gel should be destained in water to remove acids that can contribute to further denaturation of the protein. Desired antigen concentration is 0,1-1 mg/ml, however lower concentrations can also be accepted. Peptides should be in dry form unless it is conjugated with a carrier protein (such as KLH or BSA). When conjugated to carrier proteins the peptides should be sent in a buffered saline solution (no ionic detergents) with a concentration of at least 100 µg/mL. The following additives (maximum concentration) can be accepted in buffers if the concentration of the antigen is supplied at 500 µg/ml: 2 mM DTT (dithiothreitol), 1 mM 2-mercaptoethanol, 1 mM EDTA, 10% Glycerol, 200 mM Imidazole (in this case without NaCl), 0,1% Maltose, Manitol and Trehalose, 300 mM NaCl (in this case without Imidazole), 1 M Urea, 1 % Triton 100, 1 % Tween 20, 1 % b-octylglucoside, 0,1 % SDS.
Additives are toxic to animals, e.g. protease inhibitors such as 1 ug/ml Leupeptin, 0.1 mM PMSF, 0.1 mM DIPF, 1 ug/ml Pepstatin A, sodium azide should be avoided.