The Internal Protein Sequencing Research Subcom-mittee of the Protein Sequence Research Committee is conducting a study involving trypsin digestion of an SDS-PAGE-separated protein and HPLC separation of the resulting peptides. The primary goals of this study are: (i) to provide members with a mechanism to independently and anonymously evaluate their ability to generate and isolate peptides and (ii) to introduce these technologies to laboratories that do not currently employ them. Because the internal study materials were available by request only, a letter was distributed in October, 1995 to all facility directors to identify those members who wished to participate. Participants were mailed a protein sample and control in November--in the form of either a Coomassie Blue-stained SDS gel slice or a small section of Amido Black-stained PVDF membrane--and requested to digest the protein and control with trypsin and subject the digests to HPLC. For those laboratories wishing to proceed further, a specific tryptic peptide could be isolated and subjected to mass spectrometry and amino acid sequencing. All data returned by the January deadline will be included in the study results, which will be presented at ABRF '96: Biomolecular Techniques.
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