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 l 2 N HCl/50 ml a.d.)
		15 min at 37C

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 -20C,
		and dilute in 50 ml Tris/HCl buffer)
		10 min at 37C

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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