From: Howard Ratech (ratech@aecom.yu.edu)
Date: Tue Dec 26 1995 - 11:24:32 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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