Martha

	The most common, as far as I'm aware, is to use 0.1% trypsin in a
0.1% calcium chloride solution  and adjust the pH to 7.8. It may be that the
authors of the article you read used a 1% in 1% solution, if you try this I
think you stand a good change of overdigesting the tissue.

	Good luck

	John

John Auld FIBMS MSc
Biomedical Scientist 3
Immunocytochemistry
Department of Histopathology
Royal Free Hospital
Pond Street
London NW3 2QG
Tel 020 7794 0500 ext. 6516
	   


> Date: 24 Jan 2002 14:54:35 -0600
> From: mward@wfubmc.edu (Martha Ward)
> Subject: trypsin recipe
> 
> Martha Ward writes:
> 
> I have been trying to get an antibody worked up and came upon an article
> that described the procedure.  They mention the necessity of using
> "trpysin digestion (porcine trypsin, Type II, Sigma ) in calcium
> chloride (4g/100mL) for 10-30 minutes at 37 C at pH 7.8.  I am
> confused:  Do they mean 4g of the trypsin or 4g calcium chloride?  Maybe
> I am having a bad day but if anyone can help me decipher this or give me
> recipe that they are currently using I would greatly appreciate it.
> Thanks in advance for your help!
> 
> 
>