From: ccarrawa@mednet.med.miami.edu
Date: Thu, 23 Jan 97 07:18:09 EST
Message-Id: <9700238540.AA854033846@mednet.med.miami.edu>
Subject: Re: keratin
To: Recipients of ABRF List <abrf@aecom.yu.edu>
This apparently happens very frequently. ANY buffer which has become
contaminated anywhere along the way can be a problem. I tell my
students when preparing proteins for analysis (or any purpose, for that
matter): 1) NEVER use anyone else's buffers, and 2) remake all your
own buffers before starting out.
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I just seqenced a couple of peptides from a tryptic digest of a PVDF
blot. The sequences the "customer" wanted were at about the 10-15 pmole
level. In one of the peptides a second sequence starting at about the 3
pmole level was easily read out to twelve amino acids. This second
sequence turned out to be a tryptic peptide from keratin. So my
question is where did it come from, skin? As I told the customer "I
wore gloves". I recall seeing this a couple of years ago and chalked
it up to someone's poor technique and forgot about it. I wonder how
often this happens? Has any one sniped off a chunk of skin and
processed it for internal sequencing?
Steve
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Date: 22 Jan 97 17:24:15 EST
From: Stephen.A.Bobin@Dartmouth.EDU (Stephen A. Bobin)
Subject: keratin
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