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

 
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希望納入価格 (円)
国内在庫 i
2019年10月21日15時45分 現在
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#9751S100 μL66,000
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#9751T20 μL39,000
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HISTONE H3 製品一覧 | #9751 が入っているAntibody Sampler キット一覧

感度分子量 (kDa)抗体の由来貯法
内在性17Rabbit IgG-20℃
種交差性 (社内試験済)
交差する可能性がある種 i

社内試験はしていませんが、配列が100%相同であるため反応すると推定される種

ヒト、マウス、ラット、サル、キイロショウジョウバエ、出芽酵母 アフリカツメガエル、ゼブラフィッシュ
9751 の推奨プロトコール i

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推奨プロトコールはCST社内試験の徹底的なバリデーションに基づいて作成されておりますので、正確かつ再現性の高い結果が得られます。

注:各製品に最適化されたプロトコールをリンクしています。

 

ウェスタンブロッティング (1:1000)免疫組織染色 (パラフィン) (1:200)免疫蛍光細胞染色 (IF-IC) (1:400)フローサイトメトリー (1:200)クロマチン免疫沈降 (1:50)クロマチン免疫沈降シークエンス (1:50)

下記ステップについては、データシートの右側もあわせてご参照ください。

IHC-P: 抗体希釈液 / 抗原賦活化

CST推奨プロトコールに欠かせない関連製品

特異性・感度
内在性レベルのLys4 がトリメチル化されたHistone H3 タンパク質を検出します。Lys4 がジメチル化されたHistone H3 タンパク質にも若干の交差性が見られますが、Lys4 がメチル化されていない、およびモノメチル化されたHistone H3 タンパク質とは交差しません。さらに、Lys9、Lys27、Lys36 がメチル化されたHistone H3 タンパク質やLys20 がメチル化されたHistone H4 タンパク質とも交差しません。
使用抗原
Lys4 がトリメチル化されたHistone H3 タンパク質のN末端領域 (合成ペプチド)

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社内データ

※下記の社内データは、すべて9751 の推奨プロトコールで実験した結果です。

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.

Western Blotting

Western Blotting

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

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).


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).

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.

Chromatin IP-seq

Chromatin IP-seq

Chromatin immunoprecipitations were performed with cross-linked chromatin from HCT116 cells and Tri-Methyl-Histone H3 (Lys4) (C42D8) Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across RPL30, a known target gene of H3K4me3 (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.


Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either Tri-Methyl-Histone H3 (Lys4) (C42D8) Rabbit mAb or 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.

バックグラウンド

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).

使用例
 
関連製品
1064   Tri-Methyl-Histone H3 (Lys4) Blocking Peptide
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13079   Immunohistochemistry Application Solutions Kit (Rabbit)
14166   Hematoxylin
14177   SignalStain® Mounting Medium
14746   SignalStain® Citrate Unmasking Solution (10X)
15019   Animal-Free Blocking Solution (5X)
2139   LSD1 Antibody
2184   LSD1 (C69G12) Rabbit mAb
2650   Histone H3 Antibody (ChIP Formulated)
4499   Histone H3 (D1H2) XP® Rabbit mAb
4620   Histone H3 (D2B12) XP® Rabbit mAb (ChIP Formulated)
6883   SignalFire™ ECL Reagent
7074   Anti-rabbit IgG, HRP-linked Antibody
7727   Biotinylated Protein Ladder Detection Pack
9002   SimpleChIP® Enzymatic Chromatin IP Kit (Agarose Beads)
9003   SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads)
9715   Histone H3 Antibody
9717   Histone H3 (3H1) Rabbit mAb
9723   Mono-Methyl-Histone H3 (Lys4) Antibody
9725   Di-Methyl-Histone H3 (Lys4) (C64G9) Rabbit mAb
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Illumina is a registered trademark of Illumina, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.

本製品は試験研究用です。

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

Immune Cell Signaling Pathways

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