RE: Antigen Retrieval Methods

From:Jim Manavis <>


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.


Jim Manavis
Senior Hospital Scientist
Department of Neuropathology
Institute of Medical & Veterinary Science
Adelaide, SA, 5000
Phone: 61-08-8222-3668
FAX: 61-08-8222 3204
Disclaimer: Not this little black duck!

-----Original Message-----
From:	Daniel Martinez []
Sent:	Wednesday, 8 November 2000 12:15 am
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

Dan Martinez
CNDR/University of Pennsylvania

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