Martha,

This is what I do:

I make up 0.1% calcium chloride in deionized water, then adjust the pH to
7.8 using 0.1N sodium hydroxide.  Just before use, I preheat this solution
to about 37ºC, then add 0.1gm of trypsin.  Using a stir bar, I dissolve the
enzyme.  When dissolved, I incubate at 37º for 10 to 15 minutes.

Hermina

Martha Ward wrote:

> 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!

--
Hermina Borgerink, BA, HTL, HT(ASCP)IHQ
Department of Pathology
Wake Forest University School of Medicine
Winston-Salem, N.C. 27157
PH (336) 716-1538
Fax (336) 716-1515