PXD056133

PXD056133 is an original dataset announced via ProteomeXchange.

Title	ABPP-based identification of biochanin A targeted Protein in AML-12 cells
Description	AML-12 cells were stimulated with 5 mM APAP for 3 hours, with one group treated with 20 μM Bio-A and the other left untreated. Following a 30-minute incubation, both groups received 20 μM Bio-A probe and DMSO for an additional 2 hours. Total protein was extracted, and protein concentration was quantified using the BCA assay. Pre-mixed Click reaction reagents (50 mmol/L azido-tetrazine, 1 mmol/L vitamin C sodium, and 1 mmol/L CuSO4, 100 mmol/L tris-hydroxypropyltriazolylmethylamine) were then added to the lysis buffer and allowed to react for 2 hours at room temperature. Proteins were precipitated with cold acetone at -20 °C and re-dissolved in PBS containing 1.5 % SDS. The solution was incubated with streptavidin beads for 4 hours at room temperature. The streptavidin beads were washed gently with 5 mL PBS containing 1% SDS (twice), 0.1% SDS (once), and 6 M urea (three times), followed by two washes with PBS. Proteins were digested with trypsin at room temperature. The resulting peptides were desalted on a C18 column, labeled with TMT reagents, and identified using liquid chromatography-tandem mass spectrometry (LC-MS/MS, Orbitrap Fusion Lumos, Thermo Scientific, MA, USA).
HostingRepository	iProX
AnnounceDate	2024-09-16
AnnouncementXML	Submission_2024-09-24_00:08:17.455.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Rong Wu
SpeciesList	scientific name: Mus musculus; NCBI TaxID: 10090;
ModificationList	No PTMs are included in the dataset
Instrument	Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2024-09-24 00:07:59	ID requested
⏵ 1	2024-09-24 00:08:17	announced

Dataset with its publication pending

submitter keyword: ABPP, biochanin A

Shenhai Gong
contact affiliation	Southern Medical University
contact email	gsh0526@smu.edu.cn
lab head
Rong Wu
contact affiliation	Southern Medical University
contact email	wrong1995@163.com
dataset submitter

iProX dataset URI