Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

Histone H3 (D1H2) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) #12230

No. Size Price
12230S 100 µl ( 50 tests ) ¥4,264.00 现货查询 购买询价
12230 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
F 1:50 Human,Mouse,Rat,Monkey, Endogenous Rabbit IgG
IF-IC 1:3200

Species cross-reactivity is determined by western blot.

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

Homology

Species predicted to react based on 100% sequence homology: Hamster, Chicken, D. melanogaster, Xenopus, Zebrafish, Bovine,

Specificity / Sensitivity

Histone H3 (D1H2) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) recognizes endogenous levels of total histone H3 protein. This antibody does not cross-react with other histones.

Histone H3 (D1H2) XP® Rabbit mAb 兔单抗(Alexa Fluor® 647 Conjugate)能够检测内源性组蛋白H3总蛋白水平。该抗体不会与其他组蛋白产生交叉反应。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the carboxy terminus of the human histone H3 protein.

该单克隆抗体经由合成的围绕人组蛋白H3蛋白羧基端的氨基酸肽段免疫动物而生产的。

Description

This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 647 fluorescent dye and tested in-house for direct flow cytometry and immunofluorescent analysis in human cells. The antibody is expected to exhibit the same species cross-reactivity as the unconjugated Histone H3 (D1H2) XP® Rabbit mAb #4499.

该CST抗体偶联了Alexa Fluor® 647荧光染料。并通过直标法流式细胞术在人细胞内进行内部检测。它与unconjugated Histone H3 (D1H2) XP® Rabbit mAb #4499兔单抗具有相同的种属交叉反应性。

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using Histone H3 (D1H2) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) (blue pseudocolor). Actin filaments were labeled with DY-554 phalloidin (red).采用共聚焦免疫荧光术检测Hela细胞,使用的抗体为Histone H3 (D1H2) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) (蓝色,假彩色)。肌动蛋白微丝使用DY-554 phalloidin进行标记(红色)。

Flow Cytometry

Flow Cytometry

Human whole blood was fixed, lysed, and permeabilized as per the Cell Signaling Technology Alternate Flow Protocol and stained using Histone H3 (D1H2) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate). The forward/side-scatter lymphocyte gate was applied to a histogram depicting the mean fluorescence intensity of Histone H3 (blue) versus that of concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control (Alexa Fluor® 647 Conjugate) #2985 (red).按照CST 流式替代操作方法对人全血进行固定,溶解以及透性化处理,使用Histone H3 (D1H2) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate)进行染色。采用前向角/侧向角散射光设置淋巴细胞门,以直方图来表示Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb (Alexa Fluor® 488 Conjugate) (蓝色)和与之相对的浓度匹配的Rabbit (DA1E) mAb IgG XP® Isotype Control (Alexa Fluor® 647 Conjugate) #2985 (红色) 的平均荧光强度。

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).

染色质结构调整在真核生物的转录调控中扮演着一个重要角色。核小体是染色质的基本结构单位,由DNA缠绕八个核心组蛋白 (H2A、H2B、H3、H4各2个分子)组成(1)。核心组蛋白的氨基端尾部常发生多种翻译后修饰,包括乙酰化、磷酸化、甲基化和泛素化 (2-5)。这些修饰都是针对不同刺激而产生的反应,且会直接影响染色质对转录因子的可接近性,进而影响基因表达(6)。在多数物种中,组蛋白H2B 乙酰化主要发生在Lys5, 12, 15和20位点上 (4,7)。而组蛋白H3的乙酰化则主要发生在Lys9, 14, 18, 23, 27和56位点上。在一些生物体内,H3 Lys9位点的乙酰化修饰看似对组蛋白沉积和染色质组装有决定性的作用 (2,3)。组蛋白H3 Ser10、Ser28和Thr11位点上的磷酸化修饰与有丝分裂和减数分裂期间的染色体浓缩紧密相关(8-10)。组蛋白H3 Thr3位点上的磷酸化在许多物种中是高度保守的,由激酶haspin催化完成。在哺乳动物细胞内用磷酸化特异性抗体进行免疫染色,发现有丝分裂中H3 Thr3 位点的磷酸化发生在前期,而去磷酸化则发生在后期(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

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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.

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

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

Alexa Fluor is a registered trademark of Life Technologies Corporation.

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

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