| 货号 | 7123C |
| 描述 | The PathScan® Mono-Methyl-Histone H3 (Lys4) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of histone H3 when mono-methylated at Lys4. A Mono-Methyl-Histone H3 (Lys4) Rabbit Antibody* has been coated onto the microwells. After incubation with cell lysates, mono-methyl-histone H3 (Lys4) is captured by the coated antibody. Following extensive washing, biotinylated Histone H3 Rabbit Antibody* is added to detect the 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 the absorbance for this developed color is proportional to the quantity of histone H3 mono-methylated at Lys4. * Antibodies in kit are custom formulations specific to kit. |
| 反应种属 | Human |
| 应用 | ELISA |
| 目标/特异性 | CSTs PathScan® Mono-Methyl-Histone H3 (Lys4) Sandwich ELISA Kit #7123 detects endogenous levels of histone H3 when mono-methylated at Lys4. As shown in Figure 1 using the Mono-Methyl-Histone H3 (Lys4) Sandwich ELISA Kit #7123, a high level of mono-methylation at Lys4 on histone H3 is detected in COS cells when treated with TSA. The level of total histone H3 (modified and unmodified) remains unchanged as shown by Western analysis (Figure 1). Similar results are obtained when NIH/3T3 and Jurkat cells are treated with TSA (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 2. The relationship between the protein concentration of the lysate from untreated and TSA-treated COS cells and the absorbance at 450 nm is shown. 图2：未处理和TSA处理的COS细胞裂解的蛋白浓度与450 nm的吸光度读数的关系。 | |
Figure 1. Treatment of COS cells with trichostatin A (TSA) increases the mono-methylation of Histone H3 at Lys4 detected by PathScan® Mono-Methyl-Histone H3 (Lys4) Sandwich ELISA Kit #7123. TSA treatment does not affect the level of histone H3 that is detected by Western analysis. COS cells (70-80% confluent) were treated for 16-18 hours with 0.4 μM TSA at 37ºC. Absorbance readings at 450 nm are shown in the top figure while the corresponding Western blots using Histone H3 Antibody #9715 (left panel) or Mono-Methyl-Histone H3 (Lys4) Antibody #9723 (right panel) are shown in the bottom figure. 图1：使用trichostatin A (TSA)刺激COS细胞系，使得histone H3蛋白的Lys 4位点单甲基化的水平增加，随后用PathScan® Mono-Methyl-Histone H3 (Lys4) Sandwich ELISA Kit #7123检测。通过免疫印迹分析发现TSA的处理不影响histone H3蛋白水平。COS细胞(70-80% confluent)在37ºC条件下使用0.4 μM TSA处理16-18小时。在450 nm吸光度值显示在最上图中，然而通过使用Histone H3 Antibody #9715 (左图)或Mono-Methyl-Histone H3 (Lys4) Antibody #9723 (右图)，相应的免疫印迹（western blot）显示在下图中。 |
