Natural Science Forum / Biology / Biology / January 2007

Hi all,

I work with murine heart and lung endothelial cells. I treid to carry
out the fluorescence activated cell sorting analysis to find out the
expression of some of the cell surface proteins, but the problem that I
have is that, the sample histogram resembles very similar to the
negaive control.

I would really appreciate if someone could help me trouble shoot this
problem. Also what would be a good buffer to use in this assay for
endothelial cells. I used HBSS with Calcium/ magnesium with 1% FBS.

The proteins I am trying to study are E-Selectin, ICAM-1 and -2,
VCAM-1, PECAM-1, VE-Cadherin. the antibodies that I am using are all
purified monoclonal  antibodies from Rat for mouse.

The secondary antibody is FITC conjugated made in other animal against
rat.

If someone could help me to figure out my mistakes, I would really
appreciate it.

Sincerely,

Pooja