Back to Search
Start Over
Overcoming off-targets: assessing Western blot signals for Bcnt/Cfdp1, a tentative component of the chromatin remodeling complex
- Source :
- Bioscience Reports
- Publication Year :
- 2019
-
Abstract
- The Bucentaur (BCNT) protein family is characterized by a conserved amino acid sequence at the C-terminus (BCNT-C domain) and plays an essential role in gene expression and chromosomal maintenance in yeast and Drosophila. The mammalian Bucentaur/Craniofacial developmental protein 1 (Bcnt/Cfdp1) is also a tentative component of the SNF2-related CBP activator protein (Srcap) chromatin remodeling complex, but little is known about its properties, partly because few antibodies are available to examine the endogenous protein. In this paper, we assigned the Western blot signal against the mouse Bcnt/Cfdp1 as a doublet of approximately 45 kDa using anti-Bcnt/Cfdp1 antibodies, which were generated against either of two unrelated immunogens, BCNT-C domain or mouse N-terminal peptide, and in addition, the Cfdp1 knockdown mouse ES cell line and bovine tissue were used as potential negative controls. Moreover, LC-MS/MS analysis of the corresponding doublet to the Flag-tagged mouse Bcnt/Cfdp1 that was constitutively expressed in a HEK293 cell exhibited that the upper band was much more phosphorylated than the lower band with preferential Ser phosphorylation in the WESF motif of BCNT-C domain. Western blot analysis with these evaluated antibodies indicated a preferential expression of Bcnt/Cfdp1 in the early stages of brain development of mouse and rat, which is consistent with a data file of the expression of Bcnt/Cfdp1 mRNA.
- Subjects :
- 0301 basic medicine
Male
endogenous Bcnt/Cfdp1
Placenta
Biochemistry
Molecular Bases of Health & Disease
Epitopes
Mice
0302 clinical medicine
Mass spectroscopy
Antibody Specificity
Pregnancy
Structural Biology
Gene expression
Phosphorylation
Peptide sequence
Research Articles
Post-Translational Modifications
medicine.diagnostic_test
Chemistry
Brain
Gene Expression Regulation, Developmental
Nuclear Proteins
Mouse Embryonic Stem Cells
Cell biology
Phosphoprotein
Female
Protein family
Blotting, Western
Biophysics
Chromatin remodeling
Antibodies
03 medical and health sciences
Western blot
Biochemical Techniques & Resources
antibody evaluation
Chemical Biology
medicine
Animals
Humans
Rats, Wistar
Molecular Biology
Messenger RNA
RNA sequence analysis
Cell Biology
Chromatin Assembly and Disassembly
Mice, Inbred C57BL
Molecular Weight
030104 developmental biology
HEK293 Cells
Western blot analysis
Cattle
030217 neurology & neurosurgery
Subjects
Details
- ISSN :
- 15734935
- Volume :
- 40
- Issue :
- 6
- Database :
- OpenAIRE
- Journal :
- Bioscience reports
- Accession number :
- edsair.doi.dedup.....f5422a6e7ca43823dfabe7c612ddadee