Re: Proteolytic enzymes

From:Abizar Lakdawalla <> (by way of histonet)

Hi Nima, the antigen retrieval or proteolytic digestion depends on the
particular antigen-antibody pair and how the cells/tissues have been fixed. It
is pretty hard to generalize conditions unless you know the antigen-antibody
and fixation.
About the tissue coming off, there are ways to do target retrieval that will
minimize if not eliminate problems with "floaters".

Nima Farsinejad wrote:

>         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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