Re: Liver cell revisted

lawrencp@ava.bcc.orst.edu
Mon, 18 Aug 1997 09:55:51 -0700

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