CORE--PROTEIN MICROSEQUENCING

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Description: The purpose of the Protein Microsequencing Shared Resource is to provide state-of-the-art methodology in protein sequencing and protein analysis. Protein sequencing is described as being a unique technology, requiring specific, dedicated, and specialized instrumentation. The staff includes the Core Leader, a technician, and two helpers. Sequencing is done on a Beckman-Porton Sequencer, using gas phase chemistry. The amino acid analyses are done using Pico-Tag chemistry. The protocols for in-gel and on-blot tryptic digestions were developed by the staff of the facility. Investigators can either use the protocols described on the facility?s homepage or the digestion and separation of the resulting peptide fragments on a reverse phase HPLC system. The peptides present in the HPLC fractions are analyzed on a sensitive MALDI mass spectrophotometer located in the UCLA Center for Mass Spectrometry. This instrument was acquired in 1994 as a joint effort of the Protein Microsequencing Shared Resource and the UCLA Center for Mass Spectrometry, with a major portion of the funding from the CCSG. The specific goals of the facility include providing: 1) state-of-the-art methodology in protein microsequencing to the UCLA research community; 2) sensitive amino acid analyses; 3) assistance and information to laboratory investigators about determination of protein structure; and 4) other protein services, including protein cleavages, mass analyses, and isolation of internal fragments by HPLC of gel methods. The operating policies of the facility include a determination of the priority of samples received for sequencing. The highest priority goes to members of the Cancer Center and to researchers in the Department of Biological Chemistry, which provides space for the facility, and then to other members of the UCLA community.