Revision 4
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IF-F, IF-IC, FC-FP

REACTIVITY:

H M Hm Dm

SENSITIVITY:

Endogenous

MW (kDa):

17

SOURCE:

Rabbit

UniProt ID:

#P68431

Entrez-Gene Id:

8350

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunofluorescence (Frozen) 1:400 - 1:1600
Immunofluorescence (Immunocytochemistry) 1:400 - 1:1600
Flow Cytometry (Fixed/Permeabilized) 1:200 - 1:800

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

Phospho-Histone H3 (Ser28) Antibody detects endogenous levels of histone H3 only when phosphorylated at Ser28. This antibody does not cross-react with other phosphorylated histones, including phospho-histone H3 (Ser10). Non-specific cytoplasmic staining may be observed by immunofluorescence in fixed-frozen tissues.

Species Reactivity:

Human, Mouse, Hamster, D. melanogaster

Species predicted to react based on 100% sequence homology

Rat, Chicken, Xenopus, Zebrafish, Bovine

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the amino terminus of histone H3 phosphorylated on Ser28. Antibodies are purified by protein A and peptide affinity chromatography.

Background

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 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, 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).

  1. Workman, J.L. and Kingston, R.E. (1998) Annu Rev Biochem 67, 545-79.
  2. Hansen, J.C. et al. (1998) Biochemistry 37, 17637-41.
  3. Strahl, B.D. and Allis, C.D. (2000) Nature 403, 41-5.
  4. Cheung, P. et al. (2000) Cell 103, 263-71.
  5. Bernstein, B.E. and Schreiber, S.L. (2002) Chem Biol 9, 1167-73.
  6. Jaskelioff, M. and Peterson, C.L. (2003) Nat Cell Biol 5, 395-9.
  7. Thorne, A.W. et al. (1990) Eur J Biochem 193, 701-13.
  8. Hendzel, M.J. et al. (1997) Chromosoma 106, 348-60.
  9. Goto, H. et al. (1999) J Biol Chem 274, 25543-9.
  10. Preuss, U. et al. (2003) Nucleic Acids Res 31, 878-85.
  11. Dai, J. et al. (2005) Genes Dev 19, 472-88.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IF-F: Immunofluorescence (Frozen) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Revision 4
#9713

Phospho-Histone H3 (Ser28) Antibody

Western Blotting Image 1: Phospho-Histone H3 (Ser28) Antibody Expand Image
对未处理或使用 100 ng/ml nocodazole 处理 4 小时后在分裂中期同步的 CHO 和 HeLa 细胞提取物,进行蛋白质印迹分析,随后通过有丝分裂的摆脱作用分离中期细胞。蛋白印迹用 Phospho-Histone H3 (Ser28) Antibody #9713(上)或 Histone H3 Antibody #9715(下)探测。
Immunofluorescence Image 1: Phospho-Histone H3 (Ser28) Antibody Expand Image
使用 Phospho-Histone H3 (Ser28) Antibody(绿色),对出生后 1 天的大鼠脑进行共聚焦免疫荧光分析。蓝色伪彩 = DRAQ5 (DNA 荧光染料)。
Immunofluorescence Image 1: Phospho-Histone H3 (Ser28) Antibody Expand Image
使用 Phospho-Histone H3 (Ser28) Antibody(绿色),对 C2C12 细胞进行共聚焦免疫荧光分析。肌动蛋白纤丝用 Alexa Fluor®555 phalloidin(红色)进行标记。蓝色伪彩 = DRAQ5 (DNA 荧光染料)。
Flow Cytometry Image 1: Phospho-Histone H3 (Ser28) Antibody Expand Image
与碘化丙啶(DNA 含量)相比较,使用 Phospho-Histone H3 (Ser28) Antibody 对未处理的 Jurkat 细胞进行流式细胞分析。框内细胞群是 phospho-Histone H3 阳性细胞。