Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

Phospho-Histone H3 (Ser10) (D2C8) XP® Rabbit mAb (Alexa Fluor® 594 Conjugate) #8481

No. Size Price
8481S 100 µl ( 50 tests ) ¥4,264.00 现货查询 购买询价
8481 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
IF-IC 1:50 Human,Mouse,Rat,Monkey,Zebrafish, Endogenous Rabbit IgG

Species cross-reactivity is determined by western blot.

Applications Key: IF-IC=Immunofluorescence (Immunocytochemistry),

Specificity / Sensitivity

Phospho-Histone H3 (Ser10) (D2C8) XP® Rabbit mAb (Alexa Fluor® 594 Conjugate) detects endogenous levels of histone H3 only when phosphorylated at Ser10. The antibody does not cross-react with other phosphorylated histones or with acetylated histones.

Phospho-Histone H3 (Ser10) (D2C8) XP® Rabbit mAb兔单抗 (Alexa Fluor® 594 Conjugate)能够检测仅在Ser10位点磷酸化的内源性histone H3的蛋白水平。该抗体不能与其它磷酸化的组蛋白或乙酰化的组蛋白发生交叉反应。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser10 of human histone H3.

通过人工合成人源histone H3蛋白Ser10位点磷酸化的相应的磷酸化片段去免疫动物从而制备出单克隆抗体。

Description

This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 594 fluorescent dye and tested in-house for immunofluorescent analysis in human cells. This antibody is expected to exhibit the same species cross-reactivity as the unconjugated Phospho-Histone H3 (Ser10) (D2C8) XP® Rabbit mAb #3377.

Cell Signaling Technology antibody偶联了Alexa Fluor® 594 fluorescent dye,并且直接用免疫荧光分析方法内部检测人源细胞。该抗体可与非偶联的Phospho-Histone H3 (Ser10) (D2C8) XP® Rabbit mAb #3377兔单抗有相同物种的交叉反应。

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using Phospho-Histone H3 (Ser10) (D2C8) XP® Rabbit mAb (Alexa Fluor® 594 Conjugate) (red). Actin filaments were labeled with Alexa Fluor® 488 phalloidin (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

使用Phospho-Histone H3 (Ser10) (D2C8) XP® Rabbit mAb兔单抗 (Alexa Fluor® 594 Conjugate) (红色)标记,共聚焦免疫荧光分析HeLa细胞。Alexa Fluor® 488 phalloidin标记微丝蛋白(绿色)。蓝色= DRAQ5® #4084 (DNA荧光染料)。

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 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).

染色质结构的修饰在调节真核细胞的转录中扮演着重要的角色。由DNA围绕和八聚体组蛋白(H2A,H2B,H3和H4各两个)共同组成的核小体是染色质的主要组成(1)。核小体的组蛋白氨基酸尾端进行不同的转录后修饰,包括乙酰化,磷酸化,甲基化和泛素化(2-5)。这些修饰通过不同的刺激产生并且对转录因子能否接近染色质有着直接的影响,所以也影响着基因的表达(6)。在大多数的物种中组蛋白H2B主要在Lys5,,12,,15和20位点上发生乙酰化(4,7)。组蛋白H3主要是在Lys9,14,18,23,27和56位点上发生乙酰化。在某些物种里H3上的Lys9位点发生乙酰化并应该在组蛋白沉积和染色质组装中扮演着重要的角色(2,3)。组蛋白H3上Ser10,Ser28和Thr11位点的磷酸化在有丝分裂和无丝分裂中都与染色质的缩合紧密相连(8-10)。H3的Thr3位点的磷酸化在许多物种中都是高度保守的,是由kinase haspin所催化的。在哺乳动物中用磷酸化特异性的抗体做免疫组化显示H3的Thr3位点在有丝分裂的前期发生磷酸化,后期发生去磷酸化(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.

Application References

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Protocols


For Research Use Only. Not For Use In Diagnostic Procedures.

The Alexa Fluor dye antibody conjugates in this product are sold under license from Life Technologies Corporation for research use only, except for use in combination with DNA microarrays. The Alexa Fluor® dyes (except for Alexa Fluor® 430 dye) are covered by pending and issued patents. Alexa Fluor® is a registered trademark of Molecular Probes, Inc.

DRAQ5 is a registered trademark of Biostatus Limited.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

XP is a registered trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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