[ Article crossposted from bionet.molbio.methds-reagnts ]
[ Author was ]
[ Posted on 12 Jan 1995 20:33:20 GMT ]
We are trying to detect a protein on the cell surface while
the protein is mostly found intracellularly. We are fixing
with 4% paraformaldehyde but find that there is some
permeabilization of the cells. Has anyone found washing and fixing
conditions that cause very minimal damage to the cell membrane?
Does glutaraldehyde or formaldehyde leave a more intact cell
membrane than paraformaldehyde? Should washing be done with
PBS or BSS? The cells are Chinese hamster ovary cells attached
to a slide. What is the minimum % of paraformaldehyde which can
be used in fixation and will this improve the situation?
Thanks for your input. E-mail or post here.
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Ciao,
Rich
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Richard Mandel | E-mail address: rmandel at acs.bu.edu
Boston Univ. School of Medicine | Phone No. 617-638-4512
Boston, MA 02118 | Fax No. 617-638-4085
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--
Ciao,
Rich
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Richard Mandel | E-mail address: rmandel at acs.bu.edu
Boston Univ. School of Medicine | Phone No. 617-638-4512
Boston, MA 02118 | Fax No. 617-638-4085
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