ABRF-2002ESRG, a difficult sequence: Analysis of a PVDF-bound known protein with a heterogeneous amino-terminus

Scott D. Buckel, Richard G. Cook, J. Myron Crawford, David R. Dupont, Ben J. Madden, John M. Neveu, Laurey Steinke, Joseph Fernandezh

Research output: Contribution to journalArticle

6 Scopus citations

Abstract

The ABRF-2002 Edman Sequencing Research Group (ESRG) sample (ABRF-2002ESRG) was the 14th in a yearly series designed as an education and self-evaluation tool for laboratories performing Edman sequence analysis. This year's study used a known protein with a heterogeneous amino-terminus, and thus was one of the more challenging protein samples distributed by an Association for Biomolecular Resource Facilities (ABRF) research group. The sample was originally submitted to an ESRG member's lab, and after analysis was thought to demonstrate an analytical problem that would be of interest to the general sequencing community. The protein was purified using commercially available, pre-cast sodium dodecyl sulfate-polyacrylamide gels and transferred to polyvinylidene fluoride. Protein bands were distributed to 72 members of the ABRF who requested ABRF-2002ESRG, along with a data instruction sheet and a brief survey. Participating members were requested to report observed raw data, interpret the data as they normally would for an investigator, and identify the protein using a database search. Study results from 31 responses are presented to show how labs fared with a difficult but sequenceable sample.

Original languageEnglish (US)
Pages (from-to)246-257
Number of pages12
JournalJournal of Biomolecular Techniques
Volume13
Issue number4
StatePublished - Dec 2002

Keywords

  • Complex mixture
  • Heterogeneous N-terminus
  • PVDF
  • Protein/peptide sequencing

ASJC Scopus subject areas

  • Molecular Biology

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    Buckel, S. D., Cook, R. G., Myron Crawford, J., Dupont, D. R., Madden, B. J., Neveu, J. M., Steinke, L., & Fernandezh, J. (2002). ABRF-2002ESRG, a difficult sequence: Analysis of a PVDF-bound known protein with a heterogeneous amino-terminus. Journal of Biomolecular Techniques, 13(4), 246-257.