A suggestion for a more hepatocyte-specific marker would be to use an anti-cytochromeP450 enzyme antibody. Gentest (Woburn, MA) markets some anti-rat ones that cross react very well with murine cytochromeP450 enzymes. I've never used them for flow, but would expect they would work. I have had lots of problems with hepatocytes being very clumpy. To get around this I've been able to trypsinize my cells--but....depending on what you are doing, this may not be a helpful suggestion. Also, I've had better success permeabilizing with saponin (0.05%-0.1%) and no methanol. (ie, fix 1% paraformaldehyde then 5 min saponin in PBS. Dilute all antibodies and do all washes thereafter with saponin in your buffer). Good luck! B. Paige Lawrence, Ph.D. Dept. of Agricultural Chemistry Environmental Health Sciences Center Ag & Life Sciences 1007 Oregon State University Corvallis, OR 97331-7301 541-737-2096 (ph) 541-737-0497 (fax) lawrencp@bcc.orst.edu
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