Total Digest of Proteins

[Stephen R. Lasky]

In article <2sl641$r61 at nntp2.Stanford.EDU>, hkibak at leland.Stanford.EDU
(Henrik Kibak) wrote:

> In order to determine if the molecule I am interested in is a peptide,
> I would like to determine if protease digestion will abolish activity.
> 
> I have tried digesting my crude (animal tissue) homogenate with 1 mg/ml
> pronase (24 hr, room temperature, up to 200 ml total volume), but this
> enzyme alone does not completely digest all proteins to amino acids. 
>

Did you lose activity when you digested with pronase?   

You shouldn't have to digest a peptide to amino acids to abolish its
activity.  You can also use Tripsin or Chymotrypsin to knock out activity.
In addition, you can heat inactivate many proteins as well (start at 65
degrees 15 minutes).  


 

Stephen R. Lasky, Ph.D.       Brown University/Roger Williams Medical
Center
e-mail: Stephen_Lasky at brown.edu         LandLine: 401-456-6572
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