Putative K29-linked substrates identification using sAB-K29

ABSTRACT: We have identified K29-linked ubiquitination is involved in cell cycle regulation and enriched in midbody formation. To identify the substrates of K29-linked ubiquitination in the midbody, we soughtto perform immunoprecipitation (IP) under denaturing conditions to minimize the presence of noncovalently bound proteins. we synchronized HeLa cells to telophase to maximize the abundance of the midbody and lysed the cells using 8 M urea. The lysate was further diluted in 1 M urea, followed by IP using biotinylated sAB-K29. The enriched proteins were separated on an SDS-PAGE gel and subjected to label-free quantitative mass spectrometry analysis.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell

DISEASE(S): Cervix Carcinoma

SUBMITTER: Qingyun Zheng  

LAB HEAD: Lei Liu

PROVIDER: PXD024425 | Pride | 2021-07-11

REPOSITORIES: Pride