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

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    分子量：
    17
    17
    反应种属：
    Human,Mouse,Rat,Monkey,D.melanogaster,S.cerevisiae,
    Human,Mouse,Rat,Monkey,D.melanogaster,S.cerevisiae,
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    产品介绍
    产品信息
    抗原名称
    Histone H3
    来源纯化
    使用与 Lys4 三甲基化组蛋白 H3 氨基末端相对应的合成肽对动物进行免疫接种来产生单克隆抗体。
    宿主
    Rabbit
    简单描述
    Monoclonal Antibody for studying H3 (Lys4) trimethylate. Cited in 534 publications. Validated for WB, IHC, IF, F, ChIP, ChIP, C&R, C&T. Available in 2 sizes. Highly specific and rigorously validated in-house, Tri-Methyl-Histone H3 (Lys4) (C42D8) Rabbit Monoclonal Antibody (CST #9751) is ready to ship.
    商品描述

    Product Usage Information

    For optimal ChIP and ChIP-seq results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits. The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652. The CUT&Tag dilution was determined using 100,000 cells in a total reaction volume of 100 μl.

    ApplicationDilution
    Western Blotting1:1000
    Immunohistochemistry (Paraffin)1:1000 - 1:4000
    Immunofluorescence (Immunocytochemistry)1:200 - 1:800
    Flow Cytometry (Fixed/Permeabilized)1:400 - 1:1600
    Chromatin IP1:50
    Chromatin IP-seq1:50
    CUT&RUN1:50
    CUT&Tag1:50
    分子量
    17
    内毒素水平
    非洲爪蟾蜍, 斑马鱼
    研究领域
    癌症,发育生物学与干细胞研究,表观遗传学,神经科学,
    应用
    反应种属
    Human,Mouse,Rat,Monkey,D.melanogaster,S.cerevisiae,
    预测反应种属
    Xenopus,Zebrafish
    目标/特异性

    Specificity/Sensitivity

    当在 Lys4 位点进行三甲基化时，Tri-Methyl-Histone H3 (Lys4) Antibody 可检测内源水平的组蛋白 H3。已显示该抗体与 Lys4 位点二甲基化的组蛋白 H3 具有一定的交叉反应性，但是不会与非甲基化或单甲基化的组蛋白 H3 Lys4 发生交叉反应。此外，抗体不与甲基化组蛋白 H3 Lys9、Lys27、Lys36 或甲基化组蛋白 H4 Lys20 发生交叉反应。

    物种反应性：

    人, 小鼠, 大鼠, 猴, 黑腹果蝇 , 酿酒酵母

    敏感性
    Endogenous
    背景
    背景
    核小体由四个核心组蛋白（H2A、H2B、H3 和 H4）组成，是染色质的主要组成部分。最初被认为可作为 DNA 包装的静态支架的组蛋白现在已被证明是动态蛋白，可进行多种类型的翻译后修饰，包括乙酰化、磷酸化、甲基化和泛素化 (1)。组蛋白甲基化是形成基因组的活跃和非活跃区域的主要决定因素，并且在发育期间基因组的正确编程过程中发挥关键作用 (2,3)。组蛋白 H3 (Arg2, 17, 26) 和 H4 (Arg3) 的精氨酸甲基化可促进转录激活，并由蛋白精氨酸甲基转移酶 (PRMTs) 家族介导，该家族包括辅助激活因子 PRMT1 和 CARM1 (PRMT4) (4)。与此相反，已确定更多样化的组蛋白赖氨酸甲基转移酶，除了其中某个外其余的都含有一个保守催化 SET 结构域，该结构域最初在果蝇 Su(var)3-9、zeste 增强子和 Trithorax 蛋白中得以确认。赖氨酸甲基化主要发生在组蛋白 H3 (Lys4 位点, 9, 27, 36, 79) 和 H4 (Lys20 位点) 中，并已确定与转录激活和沉默有关 (4)。这些赖氨酸残基的甲基化可协调染色质修饰酶的募集，这些酶含有甲基赖氨酸结合模块，例如克罗莫结构域 (HP1, PRC1)、PHD fingers (BPTF, ING2)、tudor 结构域 (53BP1) 和 WD-40 结构域 (WDR5) (5-8)。PADI4、LSD1、JMJD1、JMJD2 和 JHDM1 等组蛋白脱甲基酶的发现表明：甲基化是可逆的表观遗传标记物 (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.
    研究领域
    癌症,发育生物学与干细胞研究,表观遗传学,神经科学,
    翻译后修饰
    Trimethylate
    制备和贮存
    保存方式
    Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
    数据库链接
    Entrez-Gene ID
    8350
    UniProt ID
    P68431

    参考图片

    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 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。

    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 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。

    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 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.

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    货号：
    9751T
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