0.05 mg of HeLa histone preps will be included in the shipment along with the primary antibody as an appropriate positive control. For SDS-PAGE with the positive control we recommend loading 0.01 mg per lane. Epi-Plus antibody production in collaboration with Rockland Immunochemicals Inc. *Chromatin immunoprecipitation was performed on this product with a standard set of primers that amplify active gene loci (GAPDH, RPL30), inactive loci (MYOD1, AFM), and heterochromatic loci (alphaSat, SAT2). The resulting data indicated there was little to no amplification of these sites, which corresponds with the current literature. Until a more specific locus can be established and validated, we give this product validity for ChIP and it is backed by the full Novus guarantee.
When confronted with DNA damage or the lack of certain kinases like Chk1 and Aurora1, histone H3 is phosphorylated at T11, causing a reduction in acetylation of the same histone at K9 and ultimately leading to a reduction of active transcription in affected cells. When the T11 phosphorylation is combined with a typical S10 phosphorylation that is critical in chromosome condensation in metaphase, the effect on transcriptional regulation is amplified further. Phosphorylation of H3S10 is decreased by T11 phosphorylation. Thus, detection of the combined modification is critical in determining transcriptional effects, especially when contrasted against single modifications of H3.