Reply - Cyclin D1 immunostaining
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From: | Richard Cartun <Rcartun@harthosp.org> (by way of histonet) |
To: | histonet <histonet@magicnet.net> |
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Content-Type: | text/plain; charset="us-ascii" |
In my experience Cyclin D1 has been one of the more difficult (or should I
say "frustrating") proteins to demonstrate in formalin-fixed, paraffin-
embedded tissue.
I agree with Keith Miller's previous comments. We also use DAKO's high pH
target retrieval solution (10' in a steamer followed by a 10' cool-down in
a pan of running cold tap water, then 2 - 5' washes in PBS) along with
their mAb (clone DCS-6) at a dilution of 1:200 for 30' with LSAB+ detection.
We recently stained sections of a mantle cell lymphoma from another
hospital and obtained spectacular immunoreactivity. It's interesting to
note that the immunoreactivity (for a variety of antigens) on tissues from
this hospital is usually weaker that what we can obtain on our own tissue.
In addition, I haven't good luck with tissue fixed in B5.
R. Cartun
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