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My question: Is it possible to demonstrate nuclear antigens with antibody
on fixed cells using only a -20 C methanol fixation step, or is it
necessary to include a second step with, e.g., Triton X-100?
The answer is yes, it is possible, and no, you don't have to
use TX-100 after MeOH.
For reference, the following list are a few of the many reviews
that are likely to be in the library or on your book shelf. There
are many others. I'm beginning to wonder why we write them.
Bauer KB and Jacobberger JW, 1994, Analysis of Intracellular
Proteins. in Methods in Cell Biology: Flow Cytometry Vol 41,
2nd edition, eds. Z. Darzynkiewicz, H. A. Crissman,
J. P. Robinson, Academic Press, Inc., NY, pp 351-376
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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