Re: trypsin recipe
|From:||email@example.com (Hermina Borgerink)|
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.
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
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