Re: Enzyme digestions
From: | "C.M. vander Loos" <c.m.vanderloos@amc.uva.nl> |
>Hi all.
>Does anyone have buffer Recipes & concentration information for using
>the following enzymes on Formalin fixed, paraffin embedded sections?
>
>Collagenase
>Elastase
>Trypsin
>CHymotrypsin
>Pepsin
>Protein Tyrosine Phospotase
>
>Thanks in advance
*********************
Dear Luis,
I have the following recipes for pretreatment of formalin-fixed and
paraffin-embedded tissue sections:
Pepsin: 0.25% pepsin (Sigma 7000) in
50 ml 10 mM HCl (250 µl 2 N HCl/50 ml a.d.)
15 min at 37°C
Trypsin: 0.1% trypsin (Sigma 8128) in
Tris/HCl (50 mM; pH 7.8)
(or: use 2 ml of 2.5% trypsin portions stored at -20°C,
and dilute in 50 ml Tris/HCl buffer)
10 min at 37°C
Chris van der Loos
Dept. of Cardiovascular Pathology
Academical Medical Centre H0-120
Meibergdreef 9
NL-1105 AZ Amsterdam
The Netherlands
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