RE: Proteolytic enzymes (by way of histonet)

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.

		-----Original Message-----
		From:	Nima Farsinejad []
		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.


		Nima Farsinejad
		Research Specialist
		Digestive Diseases
		Emory University
		Atlanta, Georgia

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