RE: Beta amyloid stain

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From:"Smith, David" <david_smith1@merck.com> (by way of histonet)
To:histonet <histonet@magicnet.net>
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Lilith
	Is the amyloid endogenous rodent or introduced human protein?  This
will have a baring on which antibodies you can use. There are numerous
antibodies to beta amyloid peptides but some are very specific for the
length of the peptide chain and the species of origin. Alkaline Congo red
works well in mouse amyloid plaques, but only seems to label plaque cores,
thioflavin S is also good.
	Recommended monoclonal clones are for human amyloid 6F3D(DAKO
1:100), needs 5mins in formic acid prior to primary to expose antigen. 6E10
is also good for the long form of the peptide (1-42). Boehringer do an anti
amyloid precursor protein (clone 22C11, use at 2.5ug/ml with formic acid
pretreatment) which will also label deposits both diffuse and dense. Sigma
have a polyclonal to 1-40 which will recognise both rodent and human beta
amyloid.
	Hope this of help.
David
Senior Research Pharmacologist
Molecular Neuropathology
Merck Sharp & Dohme
Neuroscience Research Centre
Terlings Park
Harlow, Essex CM20 2QR
+44 (0) 1279 440 492
FAX +44 (0) 1279 440 390




> ----------
> From: 	Barry, Lilith[SMTP:Lilith.Barry@nrc.ca]
> Sent: 	24 November 1998 15:40
> To: 	Histonet
> Subject: 	Beta amyloid stain
>
>  Dear colleagues,
> Which would be the best way to visualize Beta amyloid deposition in the
> rat
> brain.  There are few histocemical methods in my  Histotechnique book and
> I
> don't know which one is the most suitable.  Or should I do
> immunohstocemistry? If so, which antibody should I use?  I have very
> little
> tissue to work with, so I cannot afford experimenting much. The tissue has
> been fixed with formaldehyde  for about  2-3days and are in PBS now.
> I would very much appreciate your help.
> Lilith
>  -----------------------------------------
>
> Lilith Ohannessian-Barry
> National Research Council
> Institute of Biological Sciences
> CANADA
> Tel;613-993-6460
> Fax;613-941-4475
> e-mail; lilith.barry@nrc.ca
>




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