I am wondering about the optimum temperature for cell staining with fluorochrome tagged antibody for Flow cytometry. Majority of the articles report staining at 4C or on ice, however, some articles, especially studies on clinical research, report staining at RT. Could anyone comment on that?
I think at 4C the antibodies are stable, however it works equally fine at RT
RKG
Please specify whether you are planning to label for surface or intracellular epitopes
I have seen many protocols that stain at RT for 30 minutes. This seemed particularly long to me and so I ended up optimizing this to 10 minutes which was adequate for surface staining I was doing.
I would recommend that you do a time course for the markers you are interested in and see at what time point you reach optimum signal. If you are staining some dim markers, optimize based for that marker or any marker that you are looking to quantify.
Good luck!
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