ABRF-ESRG'03: analysis of a PVDF-bound known protein with a homogeneous amino-terminus

• Published in: Journal of biomolecular techniques Vol. 14; no. 4; pp. 278 - 288
• Main Authors: Buckel, Scott D, Cook, Richard G, Crawford, J Myron, Denslow, Nancy, Fernandez, Joseph, Madden, Ben J, Neveu, John M
• Format: Journal Article
• Language: English
• Published: United States The Association of Biomolecular Resource Facilities 01.12.2003
• Subjects: Amino Acid Sequence; Data Collection; Databases, Factual; Electrophoresis, Polyacrylamide Gel; Polyvinyls; Protein Binding; Proteins - analysis; Proteins - chemistry; Proteins - isolation & purification; Sequence Analysis, Protein
• ISSN: 1524-0215; 1943-4731

The Association of Biomolecular Resource Facilities 2003 Edman Sequencing Research Group (ABRF-ESRG'03) sample is the 15th in a series of studies designed to allow participating members to evaluate their abilities to analyze the N-terminus of a protein or peptide using automated Edman degradation chemistry. It is a follow-up study to the ESRG'02 sample, which was a single protein with a heterogeneous N-terminus. Both the 2002 and 2003 samples were obtained from the same protein complex and were resolved by SDS-PAGE followed by electrophoretic transfer to PVDF membrane. The ABRF-ESRG'03 sample had an apparent molecular weight of 49 kDa and a single N-terminus, with initial yields of approximately 2 pmol. Participants were asked to sequence 25 residues and return their results to the ESRG for analysis along with two completed surveys and an area/pmol table for repetitive and initial yield calculations. Data for 46 responses are presented which include initial yields, repetitive yields, sequencer performance, and ability to identify the protein.