I agree with Calman--the biggest problems I find with approaches to intracellular staining are in the fix/perm steps. Those who are successful (and we have been for many cytokines and nuclear proteins) take a cutomized approach to their target. We have had success with "commercial" preps, and failures and the same is true with our "home brew" approach. Some proteins and cells like saponin, some like tween, some triton, some like shorter or longer fix/perm times etc etc etc.. It is aways critical to develop a system, with all the appropriate controls to address your particular question. While commercial approaches often yield success, there is a danger to lose sight of the rational scientific approach..after 20 years+ in research, I am more convinced than ever that there is rarely a cookbook approach! -Jonni Jonni S. Moore, Ph.D. Director of Clinical and Research Flow Cytometry University of Pennsylvania School of Medicine 203 John Morgan Bldg. Philadelphia, PA 19104-6082 Phone: 215-898-6853
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