RE: Antigen Retrieval Methods
From: | Jim Manavis <jim.manavis@imvs.sa.gov.au> |
Daniel
What solution have you used for retrieval with microwaving/boiling. In our
laboratory we perform a Citrate microwave retrieval (pH 6.0) for 10 minutes
for alpha-synuclein. You might need to try a 20 minute or even try using a
different pH solution, like DAKO TRS, or even EDTA which is again a
different pH. Possibly you could try microwave retrieval followed by Trypsin
(370C for 3 min). With the tissue being in an aldehyde for that long you
will have to consider using a much harsher retrieval regimen.
Cheers
Jim Manavis
Senior Hospital Scientist
Department of Neuropathology
Institute of Medical & Veterinary Science
Adelaide, SA, 5000
Australia
Phone: 61-08-8222-3668
FAX: 61-08-8222 3204
e.mail: jim.manavis@imvs.sa.gov.au
Disclaimer: Not this little black duck!
-----Original Message-----
From: Daniel Martinez [mailto:nativenewjersian@yahoo.com]
Sent: Wednesday, 8 November 2000 12:15 am
To: histonet@pathology.swmed.edu
Subject: Antigen Retrieval Methods
I currently have a rare Parkinson's case that I
am having trouble staining. This tissue has been
fixing in formalin for 13yrs. I am attempting to
stain this tissue with several alpha-synuclein
antibodies that we produce. To date, I have tried
formic acid, proteinase-K, microwave, and boiling
treatments. Any advice on other methods that might be
worth trying would be appreciated. Thanks for your
help
Dan Martinez
CNDR/University of Pennsylvania
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