Matt, Lewis Picker had a paper a couple of years ago using PFA/saponin fix and perm and noted an IL-6 mAb was cross reacting with a histone in the cells. They then did intracellular staining with a mAb that was generated against the specific histone and got similar results. That would suggest that yes, these techniques do permeabilize the nuclear membrane. Sorry. Calman > ---------- > From: Matthew Morrow > Sent: Wednesday, May 17, 2000 11:21 AM > To: Cytometry Mailing List > Subject: cytoplasmic staining > > > Flow Gurus... > > Just a quick question regarding cytoplasmic staining: Do > commercial preps such as BD's Cytofix/Cytoperm and Caltag's Fix and > Perm also permiabilize the nuclear membrane? I'm interested in > staining for a protein typically located within the nucleus which > relocates to the cytoplasm when its cell is infected with an > intra-cellular pathogen, so I need the nuclear membrane to remain > intact to prevent false-positive signals. Any suggestions appreciated. > > Matt > Matthew Morrow, MS, MT (ASCP) > Warren Magnuson Clinical Center > National Institutes of Health > Bethesda, MD > mmorrow@nih.gov/mmorrow@mail.cc.nih.gov > (301) 496-4879 >
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