| 货号 | 7155C |
| 描述 | CSTs PathScan® 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 (Ser10) 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. |
| 反应种属 | Human/Mouse |
| 应用 | ELISA |
| 目标/特异性 | CSTs PathScan® Phospho-Histone H3 (Ser10) Sandwich ELISA Kit detects endogenous levels of Phospho-Histone H3 (Ser10). Using this Sandwich ELISA Kit #7155, 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 the Histone H3 Antibody #9715 (Figure 1). 293 cells treated with Calyculin A show similar results (data not shown). This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species. |
| 供应商 | CST |
| 背景 | Modulation of chromatin structure plays an important role in the regulation of transcription in eukaryotes. The nucleosome, made up of DNA wound around eight core histone proteins (two each of H2A, H2B, H3, and H4), is the primary building block of chromatin (1). The amino-terminal tails of core histones undergo various post-translational 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, gene expression (6). In most species, histone H2B is primarily acetylated at Lys5, 12, 15, and 20 (4,7). Histone H3 is primarily acetylated at Lys9, 14, 18, 23, 27, and 56. Acetylation of H3 at Lys9 appears to have a dominant role in histone deposition and chromatin assembly in some organisms (2,3). Phosphorylation at Ser10, Ser28, and Thr11 of histone H3 is tightly correlated with chromosome condensation during both mitosis and meiosis (8-10). Phosphorylation at Thr3 of histone H3 is highly conserved among many species and is catalyzed by the kinase haspin. Immunostaining with phospho-specific antibodies in mammalian cells reveals mitotic phosphorylation at Thr3 of H3 in prophase and its dephosphorylation during anaphase (11). |
| 存放说明 | 4C |
| 参考文献 | Workman, J.L. and Kingston, R.E. (1998) Annu Rev Biochem 67, 545-79. Hansen, J.C. et al. (1998) Biochemistry 37, 17637-41. Strahl, B.D. and Allis, C.D. (2000) Nature 403, 41-5. Cheung, P. et al. (2000) Cell 103, 263-71. Bernstein, B.E. and Schreiber, S.L. (2002) Chem Biol 9, 1167-73. Jaskelioff, M. and Peterson, C.L. (2003) Nat Cell Biol 5, 395-9. Thorne, A.W. et al. (1990) Eur J Biochem 193, 701-13. Hendzel, M.J. et al. (1997) Chromosoma 106, 348-60. Goto, H. et al. (1999) J Biol Chem 274, 25543-9. Preuss, U. et al. (2003) Nucleic Acids Res 31, 878-85. Dai, J. et al. (2005) Genes Dev 19, 472-88. Steiner, P. et al. (2007) Clin Cancer Res 13, 1540-51. |
Figure 1: Treatment of NIH/3T3 cells with Calyculin A causes accumulation of phospho-histone H3 (Ser10), detected by Sandwich ELISA kit #7155, but does not affect the level of total histone H3 protein, detected by western analysis. OD 450 readings are shown in the top figure, while the corresponding western blot using Phospho-Histone H3 (Ser10) Antibody #9701 or Histone H3 Antibody #9715, is shown in the bottom figure. 图1：使用Calyculin A刺激NIH/3T3细胞系，使得phospho-histone H3 (Ser10)的蛋白堆积，随后用Sandwich ELISA kit #7155检测，通过免疫印迹分析发现不影响histone H3的总蛋白水平。在450 nm吸光度值显示在最上图中，使用Phospho-Histone H3 (Ser10) Antibody #9701或Histone H3 Antibody #9715，相应的免疫印迹（western blot）显示在下图中。 | |
Figure 2: Linear relationship between protein concentration of lysates from untreated and Calyculin A-treated NIH/3T3 cells and kit assay optical density readings. NIH/3T3 cells (80% confluence) were serum-starved overnight, and serum was added back for 15 minutes followed by treatment with Calyculin A (0.1 μM for 15 minutes). 图2：未处理和Calyculin A处理了 的NIH/3T3细胞裂解的蛋白浓度与试剂盒分析光密度读数的关系。NIH/3T3细胞(80% confluence)血清饥饿过夜，并且血清加回后15分钟随后给予Calyculin A (0.1 μM for 15 minutes)处理。 |
