Histone H3, phosphorylated (Ser10) Sandwich ELISA Kit, BioAssay(TM)

Detects endogenous levels of Phospho-Histone H3 (Ser10). Using this Sandwich ELISA Kit Phospho- Histone H3 (Ser10) is detected when treated with Calyculin A in NIH 3T3 cells. However, the levels of Histone H3 remains unchanged, as shown by Western analysis using Histone H3. Phospho-Histone H3 (Ser10) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Phospho-Histone H3 (Ser10) protein. A Histone H3 Antibody has been coated onto the microwells. After incubation with cell lysates, both nonphospho- and phospho-Histone H3 proteins are captured by the coated antibody. Following extensive washing, a Biotinylated Phospho-Histone H3 (Ser10) Antibody is added to detect the captured phospho-Histone H3 protein. HRP-linked Streptavidin is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of optical density for this developed color is proportional to the quantity of Phospho-Histone H3 (Ser10) protein. Modulation of chromatin structure plays an important role in the regulation of transcription in eukaryotes. The nucleosome, made up of four core histone proteins (H2A, H2B, H3 and H4), is the primary building block of chromatin (1). The amino-terminal tails of core histones undergo various posttranslational modifications, including acetylation, phosphorylation, methylation and ubiquitination (2-5). These modifications occur in response to various stimuli and have a direct effect on the accessibility of chromatin to transcription factors and, therefore, on gene expression (6). In most species, histone H2B is primarily acetylated at lysines 5, 12, 15 and 20 (4,7). Histone H3 is primarily acetylated at lysines 9, 14, 18 and 23 (2,3). Acetylation at lysine 9 appears to have a dominant role in histone deposition and chromatin assembly in some organisms 2,3). Phosphorylation at Ser10 of histone H3 is tightly correlated with chromosome condensation during both mitosis and meiosis (8). Kit Contents: H5110-12N1 Biotinylated Phospho-Histone H3 (Ser10) Detection Ab, 1x11ml H5110-12N2 HRP-Linked StreptAvidin, 1x11ml H5110-12N3 TMB Substrate, 1x11ml H5110-12N4 STOP Solution, 1x11ml H5110-12N5 20X Wash Buffer, 1x25ml H5110-12N6 Sample Diluent, 1x25ml *H5110-12N7 10X Cell Lysis Buffer, 1x15ml Storage and Stability: Store *H5110-12N87 at -20 degrees C. Store other components at 4 degrees C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial prior to removing the cap.