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PXD022746

PXD022746 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleIdentification of p47(1-170)-binding proteins
DescriptionHis-tagged p47(1-170 amino acid) (100 μg) was biotinylated using EZ-Link Sulfo-NHS-LC-biotin (Pierce), bound to immobilized Streptavidin beads (Sigma), and washed with 1 M KCl in buffer (20 mM Hepes, 1 mM MgCl2, 1 mM DTT, 0.5 % Triton X-100, 5 % glycerol, pH 7.4). Rat liver tissues were homogenized in a five-fold volume (w/v) of buffer (0.1M KCl, 30mM Tris-Cl, 1 mM MgCl2, 250 mM sucrose, protease inhibitor cocktail [Roche], pH 7.4) with a Potter-Elvehjem homogenizer using two different Teflon pestles (clearance of 0.35 and 0.05 mm). The homogenate was centrifuged at 2,000g for 15 mins, and the resulting supernatant was further centrifuged at 8,000g for 20 mins. The 8,000g-supernatant was supplied with Triton X-100 to a final concentration of 0.5 %, and was used as the rat liver extract. All these procedures were carried out at 4 ℃. The rat liver extract (530μl) was precleaned by centrifugation (20,000g for 15 mins) and incubated with the p47(1-170)-immobilized beads for 2 hrs at 4 ℃. The binding proteins were separated by SDS-PAGE and stained with Coomassie Brilliant Blue. The protein bands were excised from the gel and subjected to in-gel digestion with trypsin, followed by mass spectrometry analysis using an LCQ-Deca IT mass spectrometer (Finnigan).
HostingRepositoryjPOST
AnnounceDate2020-11-27
AnnouncementXMLSubmission_2022-09-18_02:26:07.240.xml
DigitalObjectIdentifier
ReviewLevelNon peer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterSuzune Nakayama
SpeciesList scientific name: cellular organisms; NCBI TaxID: 131567;
ModificationListS-carboxamidomethyl-L-cysteine; L-methionine sulfoxide
InstrumentLTQ Orbitrap Velos
Dataset History
RevisionDatetimeStatusChangeLog Entry
02020-11-25 20:09:32ID requested
12020-11-26 14:33:32announced
22021-02-04 17:28:28announced2021-02-04: Updated FTP location.
32022-09-18 02:26:07announced2022-09-18: Updated FTP location.
Publication List
Dataset with its publication pending
Keyword List
submitter keyword: p47, Golgi, rat liver
Contact List
Hisao Kondo
lab head
Suzune Nakayama
contact affiliationKyushu university
dataset submitter
Full Dataset Link List
jPOST dataset URI
Dataset FTP location
NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.jpostdb.org/JPST001022/