Staining nuclear antigens with antibody
Antony Bakke (bakkea@ccmail.ohsu.edu)
Tue, 14 Feb 95 10:03:36 PST
In answer to your question about p53 staining. I don't have any
direct experience, but I would suggest looking at a recent paper
in Cytometry by Mikulka and Bolton, vol 17, pages 246-257, 1994.
Also some antigens are not fixed in place by ethanol and can
disperse to other sites in the cell. The cytoplasmic staining
you are seeing may really be against p53 that has diffused into
the cytoplasm during fixation and not non-specific background.
I know that myeloperoxidase in neutrophil granules does not stay
in the granules when fixed with ethanol, but attaches to the
nuclear membrane giving a perinuclear staining. The
myeloperoxidase remains in the granules when the cells are fixed
with formalin/acetone.
CD ROM Vol 2 was produced by staff at the Purdue University Cytometry Laboratories
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If you have any comments please direct them to Dr. J. Paul Robinson, Professor & Director, PUCL,
Purdue University, West Lafayette, IN 47907. Phone:(317) 494-0757; FAX (317) 494-0517; Web http://www.cyto.purdue.edu
EMAIL robinson@flowcyt.cyto.purdue.edu
