IndraLab

Statements

MYSM1 binds Flag. 8 / 8
| 8
sparser
"In addition, as proof of principle of the co-IP approach, MYSM1 interaction with ubiquitinated histone H2A (H2Aub), a known substrate of the H2A deubiquitinase, was detectable in the anti-Flag pull-down from MYSM1-Flag transfectants, as expected ( xref C, lower panel)."
32466590
sparser
"Identification of MYSM1 Interaction Partners by Co-Immunoprecipitation-coupled Mass Spectrometry in MYSM1-Flag 293T Cells."
32466590
sparser
"Subsequently, the IP eluates from MYSM1-Flag and GFP-Flag transfected 293T cells, processed according to the workflow in xref A, were subjected to SCX-HPLC MS/MS mass spectrometry in order to identify novel MYSM1 interactions in DDR."
32466590
sparser
"In differential display analyses, relevant DNA damage-dependent MYSM1 candidate interaction partners were defined as proteins differentially enriched by at least 4-fold (log2(4) = 2) in MYSM1-Flag eluates upon etoposide vs. DMSO treatment by anti-Flag vs. IgG IP, but not present in GFP-Flag eluates ( xref A)."
32466590
sparser
"Specific potential MYSM1 interactors were classified as proteins with at least > 16-fold (log2(16) = 4) differential enrichment in MYSM1-Flag anti-Flag vs. IgG eluates, but not in the GFP-Flag eluates."
32466590
sparser
"At first, robust expression of GFP-Flag and MYSM1-Flag fusion proteins, as well as DNA damage induction by etoposide, were confirmed in comparison to endogenous MYSM1 levels in parental and DMSO-treated 293 T cells by Western blot ( xref B) and by IF analyses (not shown)."
32466590
sparser
"As an internal quality control, successful and specific co-IP of fusion proteins by anti-Flag compared with control IgG were affirmed in lysates from MYSM1-Flag transfected 293T cells vs. GFP-Flag controls, respectively, prior to sample analysis by mass spectrometry ( xref C)."
32466590
sparser
"To identify new candidate MYSM1 interaction partners and further investigate the molecular function of MYSM1 in DNA damage responses (DDR), we subsequently performed co-immunoprecipitation (co-IP) experiments in 293T cells transiently overexpressing MYSM1-Flag or GFP-Flag upon treatment with etoposide, followed by an SCX-HPLC MS/MS mass spectrometry proteomics approach [ xref , xref ]."
32466590