It is far easier to use a sandwich technique, if your primary antibody is unconjugated. This can be successfully used even with 2 and 3 color staining. Trying to conjugate an antibody is time consuming and expensive if you are buying the unlabeled antibody. You need sufficient antibody to start - at least 100 ug and 1 mg would be better. After conjugating you must remove excess FITC, unconjugated and over conjugated antibody. If you need protocols for sandwich techniques, I'd be happy to send them. Best regards, Tony Bakke bakkea@ohsu.edu
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