PXD010592

PXD010592 is an original dataset announced via ProteomeXchange.

Title	Strategies to enhance the extraction of electrophoretically separated proteins from polyacrylamide gels and their application to mass spectrometry analyses (C4PR_LIV)
Description	Polyacrylamide gel electrophoresis (PAGE) is a powerful technique for separating proteins extracted from complex biological samples. Unfortunately, the difficulty of recovering intact proteins in high yield from polyacrylamide matrices often limits further analyses. We discovered that staining proteins with Coomassie brilliant blue (CBB) immediately after electrophoresis improves extraction efficiency. Post-staining, proteins widely varying in molecular weight were recovered at high efficiency with a 10 minute procedure that did not employ a surfactant. High recoveries were also obtained from dried, archived gels. Native-PAGE separated protein complexes larger than 400 kDa were recovered in an alternative procedure that substituted the mild detergent octyl-β-D glucopyranoside for CBB. Recovered proteins retained their native structure and were amenable to structural analysis by native mass spectrometry. The established workflows facilitate in-depth proteomic and protein structure analyses as well as the purification, storage, and transport of protein samples.
HostingRepository	PRIDE
AnnounceDate	2020-07-13
AnnouncementXML	Submission_2020-07-13_03:40:03.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Philip Brownridge
SpeciesList	scientific name: Saccharomyces cerevisiae (Baker's yeast); NCBI TaxID: 4932; scientific name: Drosophila melanogaster (Fruit fly); NCBI TaxID: 7227;
ModificationList	acetylated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Fusion; Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2018-07-27 02:19:01	ID requested
⏵ 1	2020-07-13 03:40:03	announced

Takemori A, Butcher DS, Harman VM, Brownridge P, Shima K, Higo D, Ishizaki J, Hasegawa H, Suzuki J, Yamashita M, Loo JA, Loo RRO, Beynon RJ, Anderson LC, Takemori N, PEPPI-MS: Polyacrylamide-Gel-Based Prefractionation for Analysis of Intact Proteoforms and Protein Complexes by Mass Spectrometry. J Proteome Res, 19(9):3779-3791(2020) [pubmed]

curator keyword: Biomedical

submitter keyword: SDS PAGE, protein extraction

Robert J Beynon
contact affiliation	Institute of Integrative Biology, University of Liverpool, Liverpool, UK
contact email	rbeynon@liverpool.ac.uk
lab head
Philip Brownridge
contact affiliation	University of Liverpool
contact email	philipjb@liv.ac.uk
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD010592
     1. Label: PRIDE project
     2. Name: Strategies to enhance the extraction of electrophoretically separated proteins from polyacrylamide gels and their application to mass spectrometry analyses (C4PR_LIV)