| 货号 | 7238C |
| 描述 | The PathScan® Acetyl-Histone H4 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of acetylated lysines on histone H4. A Histone H4 antibody has been coated onto the microwells. After incubation with cell lysates, Histone H4 is captured by the coated antibody. Following extensive washing, an Acetylated-Lysine Rabbit mAb is added to detect the acetylated lysines on the Histone H4 protein. Anti-Rabbit IgG, HRP-Linked Antibody 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 acetylated Histone H4. |
| 反应种属 | Human/Mouse/Monkey |
| 应用 | ELISA |
| 目标/特异性 | CSTs PathScan® Acetyl-Histone H4 Sandwich ELISA Kit detects endogenous levels of acetylated histone H4. Using this Sandwich ELISA Kit #7238, acetylated lysines on Histone H4 are detected when treated with TSA in Jurkat cells. However, the levels of Histone H4 remain unchanged, as shown by Western analysis using the Histone H4 Antibody #2592 (figure 1). COS and NIH 3T3cells treated with TSA 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. |
Figure 1: Treatment of Jurkat cells with TSA causes accumulation of acetylation on Histone H4, detected by Sandwich ELISA Kit #7238, but does not affect the level of total Histone H4 protein, detected by Western analysis. OD450nm readings are shown in the top figure, while the corresponding Western blot using the Acetylated Lysine mouse mAb (Ac-K-103) #9681 (left panel) or Histone H4 Antibody #2592 (right panel), is shown in the bottom figure.图1：使用Sandwich ELISA Kit #7238检测发现TSA处理Jurkat细胞会引起组蛋白H4乙酰化水平升高。但Western 检测后发现，TSA处理不会影响组蛋白H4的总水平。450mn的吸光值见上图， Western blot检测结果见下图。Western使用的抗体是Acetylated Lysine mouse mAb (Ac-K-103) #9681 (左下图) 或Histone H4 Antibody #2592 (右下图)。 | |
Figure 2: The relationship between protein concentration of lysates from untreated and TSA treated COS cells and kit assay optical density readings. COS cells were treated with TSA (4 µM overnight). An acid extraction was performed for cell lysis in the presence of 5 mM sodium butyrate.图2：未处理和经TSA处理后COS细胞裂解物蛋白浓度与试剂盒检测光密度读数间关系如图所示。COS细胞经TSA处理（4uM 过夜）。在细胞裂解物中加入5nM丁酸钠后进行酸抽提。 |
