RE: Von Willebrand Factor Staining
|From:||"Auld, John" <John.Auld@rfh.nthames.nhs.uk>|
As you are in the UK, this may apply, if you use ICN trypsin.
Apparently the active part of trypsin< for ICC, is/was chymotrypsin, which
ICN have removed or reduced in their trypsin, to improve the quality.
Unfortunately for ICC this has had the reverse effect, that's progress for
you. We now use a half and half mixture of trypsin and chymotrypsin which
works for us. Some labs have gone over to using only chymotrypsin. This may
be why you have to use two, but I've never heard of this.
However, for Von Willebrand we use microwave heating with citrate buffer,
(antibody dilution 1/1000).
Hope this is of help.
> -----Original Message-----
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> Sent: Tuesday, July 17, 2001 6:25 AM
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> Subject: Daily Digest
> Date: 16 Jul 2001 10:39:20 -0500
> From: Garry Ashton <GAshton@picr.man.ac.uk>
> Subject: Von Willebrand Factor staining
> Dear all,
> As some of you may be aware from previous e-mails I am currently
> immunostaining FFPE sections to identify blood vessels.
> Does anybody have a good reason why on a protocol I have been shown, the
> sections are treated with 0.2% proteinase (protease I think) for 8 mins at
> 37 degrees, immediately followed after washing by the more conventional
> retrieval method using trypsin.
> Any help and advice grateful
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