PXD022746
PXD022746 is an original dataset announced via ProteomeXchange.
Dataset Summary
Title | Identification of p47(1-170)-binding proteins |
Description | His-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). |
HostingRepository | jPOST |
AnnounceDate | 2020-11-27 |
AnnouncementXML | Submission_2022-09-18_02:26:07.240.xml |
DigitalObjectIdentifier | |
ReviewLevel | Non peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Unsupported dataset by repository |
PrimarySubmitter | Suzune Nakayama |
SpeciesList | scientific name: cellular organisms; NCBI TaxID: 131567; |
ModificationList | S-carboxamidomethyl-L-cysteine; L-methionine sulfoxide |
Instrument | LTQ Orbitrap Velos |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
---|---|---|---|
0 | 2020-11-25 20:09:32 | ID requested | |
1 | 2020-11-26 14:33:32 | announced | |
2 | 2021-02-04 17:28:28 | announced | 2021-02-04: Updated FTP location. |
⏵ 3 | 2022-09-18 02:26:07 | announced | 2022-09-18: 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 affiliation | Kyushu 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/ |