| 货号 | 9849T |
| 描述 | The Phospho-Histone H3 (Mitotic Marker) Antibody Sampler Kit provides a fast and economical means of evaluating phosphorylation sites associated with mitosis on Histone H3. The kit contains enough primary and secondary antibodies to perform four Western blots. |
| 目标/特异性 | All antibodies in the Phospho-Histone H3 Antibody Sampler Kit recognize Histone H3 only when modified at the indicated site. |
| 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 | -20C |
| 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. |
Flow cytometric analysis of human peripheral blood lymphocytes using Histone H3 (D1H2) XP® Rabbit mAb (blue) compared to Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red). Anti-rabbit IgG (H+L), F(ab)2 Fragment (Alexa Fluor® 647 Conjugate) #4414 was used as a secondary antibody. | |
Western blot analysis of extracts from HeLa cells, either untreated or treated with nocodazole (0.1 mg/ml for 18 hours), using Phospho-Histone H3 (Ser10) (D2C8) XP® Rabbit mAb #3377 (upper) or Histone H3 Antibody #9715 (lower). Phospho-specificity of the antibody is shown by further treatment of the lysate with λ phosphatase. 使用Phospho-Histone H3 (Ser10) (D2C8) XP® Rabbit mAb #3377兔单抗 (上图)或Histone H3 Antibody #9715 (下图)，免疫印迹(Western blot)分析HeLa细胞中Phospho-Histone H3 (Ser10)的蛋白水平，细胞分为untreated或nocodazole-treated (0.1 mg/ml for 18 hours)。抗体的磷酸化特异性通过λ phosphatase处理的裂解物进一步证明。 | |
Western blot analysis of extracts from various cell lines using Histone H3 (D1H2) XP® Rabbit mAb #4499. 使用Histone H3 (D1H2) XP® Rabbit mAb #4499兔单抗，免疫印迹(Western blot)分析不同细胞中Histone H3 (D1H2)的蛋白水平。 | |
Confocal immunofluorescent analysis of HeLa cells using Histone H3 (D1H2) XP® Rabbit mAb (green) and β-Tubulin (9F3) Rabbit mAb (Alexa Fluor® 555 Conjugate) #2116 (red). | |
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Histone H3 (D1H2) XP® Rabbit mAb. | |
Western blot analysis of extracts from various cell lines using Histone H3 (D1H2) XP® Rabbit mAb. | |
Western blot analysis of extracts from HeLa cells, either untreated or treated with nocodazole (100 ng/ml for 18 hours), using Phospho-Histone H3 (Ser10) (D2C8) XP® Rabbit mAb #3377 (upper) or Histone H3 Antibody #9715 (lower). Phospho-specificity of the antibody is shown by further treatment of the lysate with λ phosphatase. | |
Confocal immunofluorescent analysis of HeLa cells using Phospho-Histone H3 (Ser10) (D2C8) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). | |
Flow cytometric analysis of Jurkat cells using Phospho-Histone H3 (Ser10) (D2C8) XP® Rabbit mAb versus propidium iodide (DNA content). The boxed population indicates Phospho-Histone H3 (Ser10) positive cells. | |
Western blot analysis of extracts from NIH/3T3 and HeLa cells, untreated or treated with calyculin A and 20% FCS, using Phospho-Histone H3 (Thr3) Antibody #9714 (upper) or Histone H3 Antibody #9715 (lower). 使用Phospho-Histone H3 (Thr3) Antibody #9714 (上图)或Histone H3 Antibody #9715 (下图)，免疫印迹(Western blot)分析NIH/3T3和HeLa细胞，细胞分为未处理或处理了calyculin A和20% FCS。 | |
Western blot analysis of extracts from CHO and HeLa cells, untreated or synchronized in metaphase by treatment with 0.1 mg/ml nocodazole for 4 h, followed by isolation of metaphase cells by mitotic shake-off. Blots were probed with Phospho-Histone H3 (Ser28) Antibody #9713 (upper) or Histone H3 Antibody #9715 (lower). 使用Phospho-Histone H3 (Ser28) Antibody #9713 (上图)或Histone H3 Antibody #9715 (下图)，免疫印迹(Western blot)分析CHO和HeLa细胞中Phospho-Histone H3 (Ser28)和Histone H3的蛋白水平，未处理或有丝分裂中期使用0.1 mg/ml nocodazole处理4小时和随后通过有丝分裂摆脱法分离有丝分裂中期细胞。 | |
Western blot analysis of extracts from HeLa, C6 and NIH/3T3 cells, untreated or treated with nocodazole for 24 hours in the presence or absence of λ phosphatase, using Phospho-Histone H3 (Thr11) Antibody #9764 (upper) or Histone H3 Antibody #9715 (lower). 使用Phospho-Histone H3 (Thr11) Antibody #9764 (上图)或Histone H3 Antibody #9715 (下图)，免疫印迹(Western blot)分析HeLa、C6和NIH/3T3细胞中Phospho-Histone H3 (Thr11)的蛋白水平，细胞分为untreated或nocodazole-treated 24小时，加入或不加入λ phosphatase来处理。 | |
Confocal immunofluorescent analysis of C2C12 cells using Phospho-Histone H3 (Ser28) Antibody (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye). | |
Confocal immunofluorescent analysis of postnatal day 1 rat brain using Phospho-Histone H3 (Ser28) Antibody (green). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye). | |
Flow cytometric analysis of untreated Jurkat cells, using Phospho-Histone H3 (Ser28) Antibody versus propidium iodide (DNA content). The box indicates phospho-Histone H3 positive cells. |
