Dear Chuxin, actually, there is no need to use chemiluminesce. Probably much too sensitive in this case, and very tricky to record, as the signal is not linear with time, when you pipet the substrate manually. Do you have the possibility to use TMB? Do you also get high background then? What are the values you expect for positive signals - do you include a positive control? If in doubt, just coat a serial dilution of mouse IgG (or serum).
Can you be a bit more specific and describe your experiment, please? I suppose that you immunized a mouse an want to check if it was successful. before starting monoclonal production. Is your antibody against a protein or some sort of small molecule? Please also post some details about buffers, blocking procedure, washing steps etc. Wo _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
