FACS-antibody titration - (Jun/11/2008 )

Hi,

can some one please advice me on how to titrate antibodies for FACS.
My primary antibody concentration is 50ug/ml (according to the manufactuerer) and I will stain 1ml of single cell suspension uisng FACS. I need to deermine the cncentration for staiing and I've read one way of doing it is by antibody titration. I'm weak at maths, and would greatly appreciate some tips.

thanks
SF

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try several dilutions of your antibody (1/100 ; 1/200 ; 1/400 ; ...) to find the highest dilution that gives you a nice staining

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Thanks.

I'll be staining CD44 on a human cancer cell line. Can someone suggest me a positive control? I was thinking of beta-1 integrin?

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1 ml of cell suspension or 1 million?

Do them in serial dilution, have a look at the histogram and/or plot a graph and choose the best. This might be helpful http://www.microbiology.emory.edu/altman/f...itering_Abs.htm

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