Intracellular staining
Howard Ratech (ratech@aecom.yu.edu)
Tue, 26 Dec 1995 11:24:32 -0500 (EST)
We are trying to determine if COS cells are expressing a particular
protein after transient transfection. The protein is designed to be
secreted but I am hoping we can find enough of it in the cytoplasm to tell
us the experiment worked before we do a Western blot on the culture
supernatant.We have directly PERCP conjugated mouse monoclonal antibody to
this protein. Do you have any recommended protocol for permeablization,
staining, washing etc.? Since these cells are quite large, is there
anything we should be doing with the FSC and SSC, other than using linear
amplification? We are using a BD FACSCAN with LYSIS software.
Happy Holidays.
Howard
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