Cell Signaling Technology

Product Pathways - Chromatin Regulation / Epigenetics

Tri-Methyl-Histone H3 (Lys4) (C42D8) Rabbit mAb #9751

chip   chip-on-chip   chip-validated   chipkit   chromatin ip   H3.1   H3.2   H3K4   H3K4me3   Histone   Histone h3   immunoprecipitation  

No. Size Price
9751S 100 µl ( 10 western blots ) ¥3,900.00 现货查询 购买询价
9751T 20 µl ( 2 western blots ) ¥1,500.00 现货查询 购买询价
9751 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Monkey,D. melanogaster,S. cerevisiae, Endogenous 17 Rabbit IgG
IHC-P 1:200
F 1:200
IF-IC 1:400
ChIP 1:50

Species cross-reactivity is determined by western blot.

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

Homology

Species predicted to react based on 100% sequence homology: Xenopus, Zebrafish,

Specificity / Sensitivity

Tri-Methyl-Histone H3 (Lys4) Antibody detects endogenous levels of histone H3 when tri-methylated on Lys4. This antibody shows some cross-reactivity with histone H3 that is di-methylated on Lys4, but does not cross-react with non-methylated or mono-methylated histone H3 Lys4. In addition, the antibody does not cross-react with methylated histone H3 Lys9, Lys27, Lys36 or methylated histone H4 Lys20.

Tri-Methyl-Histone H3 (Lys4) Antibody检测仅在Lys4位点三甲基化的内源性histone H3。该抗体显示与Lys4位点双甲基化的histone H3具有同样的反应活性,但是不能与Lys4位点非甲基化或单甲基化histone H3发生交叉反应。另外,该抗体不与Lys9、Lys27、Lys36位点甲基化histone H3蛋白或Lys20位点甲基化的histone H4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 Lys4 is tri-methylated.

通过人工合成仅在lys4位点三甲基化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。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Tri-Methyl Histone H3 (Lys4) (C42D8) Rabbit mAb.

使用Tri-Methyl Histone H3 (Lys4) (C42D8) Rabbit mAb,免疫组化分析人源乳腺癌组织石蜡切片。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Tri-Methyl Histone H3 (Lys4) (C42D8) Rabbit mAb.

使用Tri-Methyl Histone H3 (Lys4) (C42D8) Rabbit mAb,免疫组化分析人源结肠癌组织石蜡切片。

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using Tri-Methyl-Histone H3 (Lys4) (C42D8) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).

使用Tri-Methyl-Histone H3 (Lys4) (C42D8) Rabbit mAb (绿色),共聚焦免疫荧光分析HeLa细胞。Alexa Fluor® 555 phalloidin标记微丝蛋白(红色)。

Western Blotting

Western Blotting

Western blot analysis of various cell types using Tri-Methyl Histone H3 (Lys4) (C42D8) Rabbit mAb.

使用Tri-Methyl Histone H3 (Lys4) (C42D8) Rabbit mAb,免疫印迹(Western blot)分析不同细胞中Tri-Methyl-Histone H3 (Lys4)的蛋白水平。

Western Blotting

Western Blotting

Antibody specificity was determined by Western blotting. HeLa and NIH/3T3 cell lysates were probed with Tri-Methyl Histone H3 (Lys4) (C42D8) Rabbit mAb (Panel A) or Tri-Methyl Histone H3 (Lys4) Rabbit mAb pre-adsorbed with 1.5 μM of various competitor peptides (Panels B-M). As shown, only the tri-methyl histone H3 (Lys4) peptide competed away binding of the antibody.

抗体特异性通过免疫印迹(Western blot)确定。HeLa和NIH/3T3细胞提取物的探针使用只有Tri-Methyl Histone H3 (Lys4) (C42D8) Rabbit mAb (图A)或Tri-Methyl Histone H3 (Lys4) Rabbit mAb,并且其提前孵育了1.5 μM various competitor peptides (图B-I)。如图所示,仅tri-methyl histone H3 (Lys4) peptide竞争离开该抗体的结合。

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HeLa cells and either 10 μl of Tri-Methyl-Histone H3 (Lys4) (C42D8) 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 Tri-Methyl-Histone H3 (Lys4) (C42D8) 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,相当于一。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon using Tri-Methyl-Histone H3 (K4) (C42D8) Rabbit mAb in the presence of non-methyl peptide (left) or K4 tri-methyl peptide (right).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of human whole blood cells using Tri-Methyl-Histone H3 (Lys4) (C42D8) Rabbit mAb (blue) and Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) #4414 was used as a secondary antibody. Analysis was performed on cells in the lymphocyte gate.

Background

The nucleosome, made up of four core histone proteins (H2A, H2B, H3 and H4), is the primary building block of chromatin. Originally thought to function as a static scaffold for DNA packaging, histones have now been shown to be dynamic proteins, undergoing multiple types of post-translational modifications, including acetylation, phosphorylation, methylation, and ubiquitination (1). Histone methylation is a major determinant for the formation of active and inactive regions of the genome and is crucial for the proper programming of the genome during development (2,3). Arginine methylation of histones H3 (Arg2, 17, 26) and H4 (Arg3) promotes transcriptional activation and is mediated by a family of protein arginine methyltransferases (PRMTs), including the co-activators PRMT1 and CARM1 (PRMT4) (4). In contrast, a more diverse set of histone lysine methyltransferases has been identified, all but one of which contain a conserved catalytic SET domain originally identified in the Drosophila Su(var)3-9, Enhancer of zeste, and Trithorax proteins. Lysine methylation occurs primarily on histones H3 (Lys4, 9, 27, 36, 79) and H4 (Lys20) and has been implicated in both transcriptional activation and silencing (4). Methylation of these lysine residues coordinates the recruitment of chromatin modifying enzymes containing methyl-lysine binding modules such as chromodomains (HP1, PRC1), PHD fingers (BPTF, ING2), tudor domains (53BP1), and WD-40 domains (WDR5) (5-8). The discovery of histone demethylases such as PADI4, LSD1, JMJD1, JMJD2, and JHDM1 has shown that methylation is a reversible epigenetic marker (9).

核小体是由四种组蛋白(H2A、H2B、H3和H4)组成,它是染色质的主要构成模块。起初被认为作为一个DNA包装的静态支架,现在则显示组蛋白是动态蛋白,经历多种翻译后修饰的形式,包括乙酰化、磷酸化、甲基化和泛素化(1)。组蛋白甲基化对于该基因组的活化和未活化区域的形成有着主要决定作用,并且在发育期间对该基因组的正确规划起着关键作用(2,3)。histones H3 (Arg2、17、26)和H4 (Arg3)的精氨酸甲基化促进转录调控以及通过蛋白质精氨酸甲基转移酶(PRMTs)家族的介导,包括共激活因子PRMT1和CARM1 (PRMT4) (4)。相反,多种多样的组蛋白赖氨酸甲基转移酶已经被鉴定,除了这个之外其它的都包含一个保守的催化SET区域,这个起初被鉴定在Drosophila Su(var)3-9、zeste增强子和Trithorax蛋白。赖氨酸甲基化主要发生在histones H3 (Lys4、9、27、36、79)和H4 (Lys20),并且已经涉及到转录激活和沉默(4)。这些赖氨酸残基的甲基化协调染色质修饰酶的招募包括methyl-lysine结合模块例如chromodomains (HP1, PRC1)、PHD fingers (BPTF, ING2)、tudor domains (53BP1)和WD-40 domains (WDR5) (5-8)。组蛋白例如PADI4、LSD1、JMJD1、JMJD2和JHDM1的发现已经显示甲基化是一个可逆的后生标记物(9)。

  1. Peterson, C.L. and Laniel, M.A. (2004) Curr Biol 14, R546-51.
  2. Kubicek, S. et al. (2006) Ernst Schering Res Found Workshop , 1-27.
  3. Lin, W. and Dent, S.Y. (2006) Curr Opin Genet Dev 16, 137-42.
  4. Lee, D.Y. et al. (2005) Endocr Rev 26, 147-70.
  5. Daniel, J.A. et al. (2005) Cell Cycle 4, 919-26.
  6. Shi, X. et al. (2006) Nature 442, 96-9.
  7. Wysocka, J. et al. (2006) Nature 442, 86-90.
  8. Wysocka, J. et al. (2005) Cell 121, 859-72.
  9. Trojer, P. and Reinberg, D. (2006) Cell 125, 213-7.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Protocols

Companion Products


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

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

Cell Signaling Technology is a 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.

用户评论 --- 共 0

该产品暂无评论!

我要参与评论 :

如要参与评论请先登录网站

还没有网站账户?去注册一下吧

Products

 

Applications