Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

Acetyl- and Phospho-Histone H3 (Lys9/Ser10) Blocking Peptide #1010

Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype

Species cross-reactivity is determined by western blot.

Applications Key:

Description

This peptide is used to block Acetyl- and Phospho-Histone H3 (Lys9/Ser10) Antibody #9711 reactivity.

该多肽产品适用于阻断Acetyl-和Phospho-Histone H3 (Lys9/Ser10) Antibody #9711的活性。

Quality Control

The quality of the peptide was evaluated by reversed-phase HPLC and by mass spectrometry. The peptide blocks Acetyl- and Phospho-Histone H3 (Lys9/Ser10) Antibody #9711 by immunohistochemistry and Western Blotting.

通过反向液相色谱RP-HPLC和质谱对肽段进行分析。通过免疫组化(immunohistochemistry)和免疫印迹(Western blot)检测,该肽段阻断 Acetyl-和Phospho-Histone H3 (Lys9/Ser10) Antibody #9711。

Western Blotting

Western Blotting

Western blot analysis of Acetyl- and Phospho-Histone H3 (Lys9/Ser10) in control and serum, calyculin A and TSA treated NIH/3T3 cell extracts of Acetyl- and Phospho-Histone H3 (Lys9/Ser10) #9711 alone (left) and the same antibody preincubated with Acetyl- and Phospho-Histone H3 (Lys9/Ser10) Blocking Peptide (right).

免疫印迹(Western blot)分析NIH/3T3细胞中Acetyl- and Phospho-Histone H3 (Lys9/Ser10)蛋白水平,细胞分为对照和serum, calyculin A and TSA处理,使用Acetyl- and Phospho-Histone H3 (Lys9/Ser10) #9711 alone (左图)和预先孵育了Acetyl- and Phospho-Histone H3 (Lys9/Ser10) Blocking Peptide的同样抗体(右图)。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical staining of paraffin-embedded human breast carcinoma, using Acetyl- and Phospho-Histone H3 (Lys9/Ser10) Antibody #9711 preincubated with irrelevant control peptide (left) and Acetyl- and Phospho-Histone H3 (Lys9/Ser10) Blocking Peptide (right).

使用Acetyl- and Phospho-Histone H3 (Lys9/Ser10) Antibody #9711,其分别预先和不相关的对照多肽 (left)和Acetyl- and Phospho-Histone H3 (Lys9/Ser10) Blocking Peptide(右图)处理孵育,免疫组化(immunohistochemistry)染色人类乳腺癌组织的石蜡切片。

Applications

Use as a blocking reagent to evaluate the specificity of antibody reactivity in immunohistochemistry and Western blot protocols.

在免疫组化(immunohistochemistry)和免疫印迹(Western blot)实验中,使用该阻断肽段来检测抗体反应的特异性。

Directions for Use

For immunohistochemistry, add twice the volume of peptide as volume of antibody used in 100 µl total volume. Incubate for a minimum of 30 minutes prior to adding the entire volume to the slide. Recommended antibody dilutions can be found on the relevant product data sheet. 
 For Western immunoblotting, add 10 µl of antibody and 10 µl of blocking peptide to 10 ml of antibody dilution buffer, and incubate at room 
 temperature for 30 minutes before allowing to react with the blot.

在免疫组织化学(immunohistochemistry)中,在100µl体系加入两倍抗体体积的该试剂。并且在将总反应体积加到切片至少30 minutes之前开始孵育多肽。抗体的推荐稀释度标注在产品说明书上。对于免疫印迹(Western blot),在10ml稀释液中加入10µl抗体和10µl 阻断肽段,且在进行免疫印迹实验30 minutes之前开始室温孵育多肽。

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.

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

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

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