Dataset Information

Overcoming off-targets: Assessing western blot signals for Bcnt/Cfdp1, a tentative component of the chromatin remodeling complex

ABSTRACT: The mammalian Bucentaur/Craniofacial developmental protein 1 (Bcnt/Cfdp1) is a tentative component of the Srcap (SNF2-Related CBP Activator Protein) chromatin remodeling complex, but little is known about its properties, partly because few antibodies are available to detect the endogenous protein.　We evaluated anti-Bcnt/Cfdp1 antibodies, which were generated against either of two unrelated immuno-gens, BCNT-C domain, or mouse N-terminal peptide. As a result, we assigned the signal of the endogenous mBcnt/Cfdp1 as a doublet band of ~45-kDa using western blot analysis. LC-MS/MS analysis of the corresponding doublet to the Flag-tagged mouse Bcnt/Cfdp1 exhibited that the upper band was much more phosphorylated than the lower band having preferential Ser phosphorylation in the DWESF motif in the BCNT-C domain. These features are very similar to those of His-tagged hBCNT/CFDP1, as previously reported(Biosci Rep. 2015; 35, pii: e00228.)

INSTRUMENT(S):

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Cell Culture

SUBMITTER: Takehiro Suzuki