Nuclear biomarkers

Becky Bonner (Becky.Bonner@CCLINK.NET.uokhsc.edu)
Mon, 17 Jul 95 16:46:17 CST

We are interested in developing some assays for nuclear biomarkers in
cancer cells. The problem we are having is that we need to strip off
the cytoplasm for this particular marker since the biomarkers we are
interested in is also located in the cytoplasm...and we want to
quantify the nuclear amount. We had no problem in tissue culture
cells but did run into a problem with normal cells being resistent to
the procedure. We need a gentle method to "disaggregate" nuclei that
will leave nuclear antigens in tact. The cancer cells (from both cell
lines and human tumors) seem to work OK since their membranes are more
fragile. Sonification would seem to me to select for the most normal
cells in a tumor sample even though it might work.

I was thinking that maybe some modification of Hedley's technique to
disaggretate solid tumors might work. Need some advice and/or ideas.
Anyone doing this sort of thing? Even failures are welcomed - since
it would indicate what we shouldn't bother trying. :-)

thanks in advance for your help

Becky. :-)
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Becky Bonner
Quanititative Fluorescent Image Analysis Laboratories
Dept. of Urology, BMSB 140
University of Oklahoma Health Science Center
940 SL Young
Oklahoma City, OK 73104 USA
TEL: (405) 271-6498
FAX: (405) 271-3118
e-mail: becky-bonner@uokhsc.edu
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