Re: [Histonet] Paraformaldehyde fixation and membrane Antibody

From:"Mark Tarango"

I'd worry about "wrong staining" when you see something that looks funny.
Have you already tried staining?  You might have the best luck purchasing a
commercial reagent for this, like cytofix/cytoperm from BD.  Lots of labs
(both clinical and research) use this stuff.

Determining the best fixative often depends on the antigen of
interest.  I'm assuming these cells are from culture...  Are they suspension
or are they adherent cells?  If they are adherent cells, are you going to
trypsanize and stain them in suspension?  Are you using something like
chamber slides?

2008/3/5 karen Cai

> Hello All,
> I am just wondering whether it's OK to use 4% paraformaldehyde as
> fixation agent for cells when I try to detect the membrane protein.
> Which kind of fixation agent is the best? I know formaldehyde is a
> cell-permeable agent. So even I don't use the Triton X-100 to
> permeablize the cell, the reagent can still move readily into living
> cells and cause wrong staining. Am I right?
> Thanks in advance,
> Best Regards,
> Karen
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