PXD048481-1

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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