Re: H-Ras

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From:Michael Stumm <stumm@ubaclu.unibas.ch>
To:"Mcleod, H, Heather" <HMCLEOD@chempath.uct.ac.za>
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Date:Wed, 08 Sep 1999 14:20:23 +0200
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Dear Heather

from the Santa Cruz Catalogue I can see that this antibody is a mouse
monoclonal IgG1 provided in concentration odf 200ug/ml. Specifications
include paraffin embedded sections.
Having many antibodies from Santa Cruz and knowing many colleagues working
with them I made the experience that some of the antibodies are not working
(either on WB or IHC). After making contact with SC I was told that any
antibody is tested on acetone fixed tissue. This means immunohistochemistry
in the catalogue. Only if the antibody was tested on paraffin as well you
will see the addendum 'including paraffin sections'. Up to now SC does not
like to provide the user with pictures  of any antibody tested either on
cryo or paraffin. And although I asked for picture data sheets I never got
some to see.
What we do in the lab with these antibodies: We perform a whole test
series: either on cryosections or paraffin sections (including different
antigen retrieval methods and different dilution steps of the primary
antibody). Needs a lot of time and energy, indeed.
In your case I suggest the following procedure:
1) Ask your local SC supplier under which condition this antibody works on
paraffin and whether long fixation with formalin has a negative influence
on IHC.
2) Perform your IHC with the pretreatment needed.
3) If you don't obtain results go back to the firm and complain. Try to get
a new lot or search for another antibody that works on paraffin.

I hope this will helpyou. Maybe other histoneters can share their
experience with us.

Michael

Michael Stumm, MD, Lab 405
Department of Research
Kantonsspital Basel
Hebelstrasse 20
4031 Basel
Switzerland
phone: ++41/61/265 23 84
email: stumm@ubaclu.unibas.ch





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