RE: Proteolytic enzymes
From: | Patsy.Ruegg@UCHSC.edu (by way of histonet) |
Nima,
I do enzyme antigen retrieval exclusively as I feel it is easier to control
than heat. I mostly use proteinase k for 5 min. on paraffin sections,
should need even less for cells, or pepsin for 10 min. at 37 dC on paraffin
sections, again you should need less with cells.
Patsy
-----Original Message-----
From: Nima Farsinejad [mailto:nfarsin@emory.edu]
Sent: Thursday, August 31, 2000 12:46 PM
Cc: Histonet
Subject: Proteolytic enzymes
I have noticed that most antigen retrieval protocols
that were
sent to me (and many thanks to everyone that did!) included
some type of
heat treatment. However I want to perform single cell gene
array studies
and I do not wish to heat the tissue, therefore I am
thinking about using
a proteolytic enzyme instead (Proteinase K, Pepsin, pronase,
or trypsin).
However, before I do this, I want to ask the very smart
people in Histonet
:o) about your opinion on using this enzymes and the
possibility of some
other technique that might compensate the antigen masking
produced by
formaldehyde fixation of paraffin embedded tissue.
Thanks,
Nima Farsinejad
Research Specialist
Digestive Diseases
Emory University
Atlanta, Georgia
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