Asxl2 is a chromatin factor that regulates histone H3 methylation and H2A deubiquitination. Previous research in our lab has shown that Asxl2... Show moreAsxl2 is a chromatin factor that regulates histone H3 methylation and H2A deubiquitination. Previous research in our lab has shown that Asxl2 is an important regulator of mammalian heart development and function. To facilitate the characterization of Asxl2, we proposed to generate an affinity reagent for the detection of Asxl2 proteins in vivo and in vitro. A recombinant GST-Asxl2 fusion protein in E. coli was produced that contained GST and the N-terminal 390 amino acids of Asxl2, separated by a Precision protease cleavage site. Asxl2 N1-390 was purified from GSTAsxl2 N1-390 and the purified polypeptide was biotinylated and used to select a phagedisplay antibody library. Finally, six antibodies were produced that exhibit various degrees of affinity with the recombinant Asxl2 N1-390 in ELISA assay. In the future the antibodies will be tested for their ability to detect endogenous Asxl2 proteins in western blot assay. This would facilitate the study of the role of Asxl2 in heart development and function. M.S. in Biology, May 2013 Show less
