Cyanogen Bromide Digest Question

D.M. Bunk dmb618 at enh.nist.gov
Fri Mar 31 10:20:32 EST 1995


I am planning on doing a cyanogen bromide digest of 
huamn serum albumin next week and I was looking for 
some pointers from anyone with CNBr digest experience.

1.	The method I found calls for dissolving the 
protein in 70% formic acid, adding solid CNBr (2 mg 
CNBr/mg protein), and then letting react for 24h at RT 
in the dark.  Is this a reliable and efficient 
procedure?

2.	If I try the CNBr digest on HSA without first 
reducing the disulfides, will the CNBr cleave or 
modify the disulfides?

3.	If I do reduce the disulfides of HSA but do 
not alkylate before the CNBr digest, will the 
cysteines end up alkylated (to the isothiocyanate)?

Any help would be appreciated!

-David




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