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Proteins can be fixed inside a cell because they can be 1.)
cross-linked or 2.) made insoluble by the fixation process; this
latter process has been termed coagulative fixation and is due to
partial denaturation. A small organic molecule like a drug, is
unlikely to be able to be cross linked by PFA and will not denature as
a protein would. If you got lucky and by chance did have a drug that
had a site that was amenable to cross linking, it would likely disrupt
the Ab binding site. Proteins work in part because they are large
molecules and you can fiddle with one part without disrupting the Ab
binding site.
Call me Mr. Optimist!
Calman Prussin
Laboratory of Allergic Diseases
NIAID/ National Institutes of Health
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From: Richard K. Meister
Sent: Wednesday, May 28, 1997 3:20 PM
To: cyto-inbox
Subject: Drug studies
Hi, everyone:
I have an investigator who wants to look at intracellular distribution
of a
drug using confocal laser scanning microscopy. The drug is not
fluorescent,
so he has a monoclonal antibody (FITC conjugate) to the drug. Our
question
is, during permeablization to allow the antibody into the cells, what
is the
best way to prevent the drug from leaching out of the cells? I
suppose a
pre-permeablization paraformaldehyde fixation step would be helpful,
but I
would like to hear from anyone who has done such drug studies and
knows if
this is an effective approach.
Thanks in advance.
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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web