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In fact we hope to do two-color fluorescence: p53 together with a
cell-surface antigen.
We tried first using flow analysis (we have a FACS 440) and had some
problems with high background with second antibody only controls so then
decided to use our Meridian ACAS.
An investigator using our ACAS today told me they have had good luck
staining nuclear antigens using a two-step fixation/permeabilization. The
first step is with 1% formalin containing cacodylate (sp?) for 20 min at
room temp followed by -20 C acetone for 5 min. I think we'll give that a
try.
Thanks for the comments we have received so far especially with regard to
the fact that the cytoplasmic staining may not be non-specific but
nuclear antigen that has leaked during fixation/permeabilization.
Ray
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Raymond B. Hester "We want only to show you
Flow Cytometry Lab something we have seen and to tell
The University of South Alabama you something we have heard...
Mobile, AL 36688-0002 that here and there in the world
Email: rhester@jaguar1.usouthal.edu and now and then in ourselves
Voice: (205)460-6029 is a New Creation."
- Paul Tillich, The New Being
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