PXD048481

PXD048481 is an original dataset announced via ProteomeXchange.

Title	Proteome analysis of mouse liver at ad libitum feeding
Description	The liver samples were collected from ad libitum feeding and Lys(6)-SILAC (Silantes, München, Germany) 10-week-old wild-type and ob/ob mice. Tissues were lysed with 0.5 mL of a solution containing 50 mM Tris-HCl pH8.8, 2% sodium dodecyl sulfate (SDS), and 7 M urea, and then homogenized by ultrasonication using a Bioruptor (Digaenode, Denville, NJ, USA). After dilution with water, the samples were centrifuged at 15,000 g for 15 min at 4°C to exclude the insoluble fractions. By calibrating to 1 mg/mL, bicinchoninic acid (BCA) assay (Thermo Fisher Scientific) was used to quantify the protein concentration of the lysates. Then, 50 µg of non-labeled and labeled protein extracts were combined in another tube for peptide preparation. We blocked cysteine residues using 2 mM Tris (2-carboxyethyl) phosphine hydrochloride (TCEP) (Thermo Fisher Scientific) at 37 °C for 30 min followed by alkylation with 10 mM 2-iodoacetamide (IAA) at room temperature for 30 min. By ultrasonically treating the pellet three times for 30 s with intervals of 30 s with a Bioruptor (Diagenode), the proteins precipitated with acetone for 3 h at −30 °C were dispersed in 50 mM triethylammonium bicarbonate. Lysyl endopeptidase (Wako, Osaka, Japan) was added to the protein suspension and digested at 37 °C for 16 h. Prior to MS analysis, the resulting peptides were purified using C18-StageTip purification after centrifuging at 15,000 g for 15 min at 4°C. Before MS analysis, peptides were resolved in 50 µL 3% ACN-0.1% foromic acid.
HostingRepository	jPOST
AnnounceDate	2024-01-12
AnnouncementXML	Submission_2024-01-11_17:11:13.301.xml
DigitalObjectIdentifier
ReviewLevel	Non peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Keigo Morita
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090;
ModificationList	S-carboxamidomethyl-L-cysteine; 6x(13)C labeled L-lysine
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2024-01-11 16:56:01	ID requested
⏵ 1	2024-01-11 17:11:13	announced

Dataset with its publication pending

submitter keyword: mouse, liver, ob/ob, WT, ad libitum

Shinya Kuroda
lab head
Keigo Morita
contact affiliation	University of Tokyo
dataset submitter

jPOST dataset URI

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