Donald Rudikoff

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From:"Patterson, Noelle" <PattersonN@NMRIPO.NMRI.NNMC.NAVY.MIL> (by way of histonet)
To:histonet <>
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I tried to send this direct.  But, it got kicked back as user address
unknown.  Are you sure you have the letter case correct?  I think it may
make a difference to some servers and not others.

Noelle Patterson
Naval Medical Research Center
Bethesda, Md

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From:	"Patterson, Noelle" <PattersonN@NMRIPO.NMRI.NNMC.NAVY.MIL
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To: <>
Subject:	RE: Thank you everyone
Date:	Fri, 29 Jan 1999 09:46:14 -0500
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I have done this in monkeys. We use antibodies to human B7-1 and B7-2.  They
stain fairly pail, but at reasonable concentrations.  We are using 1F1 and
3D1 antibodies from Genetics Institute.  We have a CRADA with them, so I
don't know if they are ready for retail sale yet.  I have tried pharmingen
antibody for this, and it did not work convincingly.
I have stained lymphocyte surface molecules in lymph node and kidney...but
have not yet tried in skin.  10 minutes in -20C acetone works well.  I then
dip 10 times in water, 10 times back in the acetone (this is to wash away
excess OCT, then quick dry the slides again).  When running on the automated
stainer I let air dry 1 min, and then do another 10-15 dips in the same
acetone.  This "extra fixation" helps to keep the tissue on the slides,
since they undergo more force from the automated stainers washing steps.
When on the manual stainer system, this extra fixation step is not
necessary.  I have tried skin using the same protocol once, poor results due
to improperly stored and prepared tissue. I haven't gotten back to it.
Good luck.  I would be happy to answer further questions, but am going out
of town for a week, starting this afternoon.  Feel free to email me, it just
may be a week and a few days before I get back to you.
Again, this is in monkeys with human-specific antibodies and may be a little
different.  But, should work!
Noelle Patterson
Naval Medical Research Center
Bethesda, Md

	From: <>
[] <mailto:[]>
	Sent:	Thursday, January 28, 1999 10:15 PM
	Subject:	Thank you everyone

Just wanted to thank everyone who responded to my questions
regarding fixation
and incubation.  Another question: Is there an optimum time and
for fixing frozen human skin sections in acetone for detection of
cell surface markers as well as IgE?
Second Question: Has anyone stained for CD80 (B7-1) and CD86 (B7-2)
and FcER1
in frozen tissue.
Any help will be appreciated,
Donald Rudikoff, MD
Dept of Dermatology, Mt Sinai School of Medicine

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