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Has anyone done a optimization on a antibody?

I need to do a optimization on a antibody and I have no idea where to start. Do you have any idea where I can find a website that would explain a procedure or have any information?

Public Comments

1. this greatly depends on your application -- western blot, ELISA, flow cytometry, etc... Whatever it may be, you need a sample that contains your antigen of interest and use your antibody as you normally would, but do multiple dilutions using just your positive control (and a negative control that lacks your antigen). that is, do ~5 10-fold dilutions of your antibody to determine how much you need to use to detect your antigen without wasting too much antibody. some applications are more sensitive that others to antibody concentration for a clear result (like a western blot). others (like flow cytometry) are less sensitive as far as results/detection go, but you dont want to waste antibody by using more than you need.