Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb #9649

chip   chip-on-chip   chip-validated   chipkit   chromatin-IP   H3.1   H3.2   immunoprecipitation   sc-8655  

No. Size Price
9649S 100 µl ( 10 western blots ) ¥3,900.00 现货查询 购买询价
9649T 20 µl ( 2 western blots ) ¥1,500.00 现货查询 购买询价
9649 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Monkey,Zebrafish,C. elegans, Endogenous 17 Rabbit IgG
IP 1:25
IHC-P 1:800
F 1:200
IF-IC 1:400
ChIP 1:50

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, IHC-P=Immunohistochemistry (Paraffin), F=Flow Cytometry, IF-IC=Immunofluorescence (Immunocytochemistry), ChIP=Chromatin IP,

Homology

Species predicted to react based on 100% sequence homology: S. cerevisiae,

Specificity / Sensitivity

Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb detects endogenous levels of histone H3 only when acetylated on Lys9. This antibody does not cross-react with other acetylated histones.

Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb兔单抗能够检测仅在Lys9位点乙酰化的内源性histone H3蛋白。该抗体不能与其它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 peptide corresponding to the amino terminus of histone H3 in which Lys9 is acetylated.

通过合成的仅在Lys9位点乙酰化的人源histone H3蛋白氨基端相应的多肽片段去免疫动物从而制备出此单克隆抗体。

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with TSA #9950 (right), using Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).

使用Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb兔单抗 (绿色),共聚焦免疫荧光分析HeLa细胞,细胞分为untreated (左图)和TSA-treated (#9950;右图)。DY-554 phalloidin标记微丝蛋白(红色)。

Western Blotting

Western Blotting

Western blot analysis of lysates from HeLa and NIH/3T3 cells, untreated or TSA-treated (400 nM for 18 hours) using Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb.

使用Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb兔单抗,免疫印迹(Western blot)分析HeLa和NIH/3T3细胞中Acetyl-Histone H3 (Lys9)的蛋白水平,细胞分为untreated或TSA-treated (400 nM for 18 hours)。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human osteosarcoma using Acetyl-Histone H3 (Lys 9) (C5B11) Rabbit mAb.

使用Acetyl-Histone H3 (Lys 9) (C5B11) Rabbit mAb兔单抗,免疫组化分析石蜡包埋的人源成骨肉瘤切片。

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 10 μl of Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human RPL30 Exon 3 Primers #7014, SimpleChIP® Human GAPDH Exon 1 Primers #5516, SimpleChIP® Human MyoD1 Exon 1 Primers #4490, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

使用SimpleChIP®Enzymatic Chromatin IP Kit (Magnetic Beads) #9003,用4 x 106 HeLa细胞的交联染色质以及10 µl Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb或2 µl Normal Rabbit IgG #2729进行染色质免疫沉淀实验。使用SimpleChIP® Human RPL30 Exon 3 Primers #7014、SimpleChIP® Human GAPDH Exon 1 Primers #5516、SimpleChIP® Human MyoD1 Exon 1 Primers #4490和SimpleChIP® Human α Satellite Repeat Primers #4486,浓缩的DNA通过real-time PCR定量。在每个样品中免疫沉淀DNA的数量被当做一个相对于总input chromatin的数量的信号,这相当于一。

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of untreated HeLa cells using Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb #9649 versus propidium iodide (DNA content). Note positive staining in cycling cells (box).

与propidium iodide (DNA含量)比较,使用Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb #9649兔单抗,流式细胞仪分析未处理的HeLa细胞。显示在细胞周期中呈阳性染色 (方框)。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human gastic carcinoma using Acetyl-Histone H3 (K9) Rabbit mAb in the presence of non-acetyl-peptide (left) or K9 acetyl-peptide (right).

使用Acetyl-Histone H3(K9) Rabbit mAb兔单抗,免疫组化分析人胃癌组织石蜡切片,左图是非乙酰化的肽段,右图是K9乙酰化的肽段。

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

U.S. Patent No. 5,675,063.

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

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