PXD015347

PXD015347 is an original dataset announced via ProteomeXchange.

Title	TMT6plex_bRP9_sperm_PITHD1_WT_KO
Description	Sperm isolated from WT mice and PITHD1-deficient mice in triplicate were lysed in 6 M guanidine-HCl containing 100 mM Tris-HCl, pH 8.0, and 2 mM DTT. The lysates were dissolved by heating and sonication, followed by centrifugation at 20,000 × g for 15 min at 4 °C. The supernatants containing 100 µg protein each were reduced in 5 mM DTT at room temperature for 30 min and alkylated in 27.5 mM iodoacetamide at room temperature for 30 min in the dark. Proteins were purified by methanol/chloroform precipitation and solubilized with 25 µL of 0.1% RapiGest SF (Waters) in 50 mM triethylammonium bicarbonate buffer. After repeated sonication and vortexing, the proteins were digested with 1 μg of trypsin/Lys-C mix (Promega) for 16 h at 37 °C. Approximately 25 µg of peptides for each sample was labeled with 0.2 mg of TMT6plex reagents for 1 h at room temperature. After the reaction was quenched with hydroxylamine, all the TMT-labeled samples were pooled, acidified with trifluoroacetic acid (TFA), and fractionated using a Pierce High pH Reversed-phase Peptide Fractionation Kit (Thermo Fisher). Nine fractions were collected using 5%, 10%, 12.5%, 15%, 17.5%, 20%, 22.5%, 25%, and 50% acetonitrile (ACN) in 0.1% triethylamine. Each fraction was evaporated in a SpeedVac concentrator and dissolved in 0.1% TFA.
HostingRepository	jPOST
AnnounceDate	2020-01-10
AnnouncementXML	Submission_2022-09-18_02:26:59.910.xml
DigitalObjectIdentifier
ReviewLevel	Non peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Hidetaka Kosako
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090;
ModificationList	S-carboxamidomethyl-L-cysteine; alpha-amino acetylated residue; L-methionine sulfoxide
Instrument	instrument

Revision	Datetime	Status	ChangeLog Entry
0	2019-09-08 21:40:40	ID requested
1	2020-01-09 18:29:13	announced
2	2021-02-04 17:06:28	announced	2021-02-04: Updated FTP location.
⏵ 3	2022-09-18 02:27:00	announced	2022-09-18: Updated FTP location.

Dataset with its publication pending

submitter keyword: TMT, sperm, PITHD1

Hidetaka Kosako
lab head
Hidetaka Kosako
contact affiliation	Tokushima University
dataset submitter

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/JPST000673/