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#4909 Tri-Methyl-Histone H3 (Lys36) (D5A7) XP® Rabbit mAb

 
CSTコード 包装
希望納入価格 (円)
国内在庫 i
2019年8月23日15時25分 現在
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#4909S100 μL72,000
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#4909T20 μL
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HISTONE H3 製品一覧 | #4909 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 ハムスター、ニワトリ、キイロショウジョウバエ、アフリカツメガエル、ゼブラフィッシュ、ウシ
4909 の推奨プロトコール i

最適な結果を得るために:Cell Signaling Technology (CST) 社は、各製品の推奨プロトコールを使用することを強くお薦めいたします。
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) (1:800)フローサイトメトリー (1:50)クロマチン免疫沈降 (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Lys36 がトリメチル化されたHistone H3 タンパク質を検出します。Lys36 がメチル化されていない、モノ、あるいはジメチル化されたHistone H3 タンパク質とは交差しません。また、Lys4、Lys9、Lys27 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
Lys36 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4909 の推奨プロトコールで実験した結果です。

ELISA

ELISA

Tri-Methyl Histone H3 (Lys36) (D5A7) XP® Rabbit mAb specificity was determined by peptide ELISA. The graph depicts the binding of the antibody to pre-coated tri-methyl histone H3 (Lys36) peptide in the presence of increasing concentrations of various competitor peptides. As shown, only the tri-methyl histone H3 (Lys36) peptide competed away binding of the antibody.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Tri-Methyl-Histone H3 (Lys36) (D5A7) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human papillary carcinoma of the breast using Tri-Methyl-Histone H3(K36) (D5A7) XP(R) Rabbit mAb.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Tri-Methyl-Histone H3(K36) (D5A7) XP(R) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded 786-O cell pellet (left, positive) or A498 cell pellet (right, negative) using Tri-Methyl-Histone H3(K36) (D5A7) XP(R) Rabbit mAb. Note that the A498 cell line harbors a SETD2 mutation.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human serous papillary carcinoma of the ovary using Tri-Methyl-Histone H3(K36) (D5A7) XP(R) Rabbit mAb in the presence of non-methyl peptide (left) or K36 tri-methyl peptide (right).


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using Tri-Methyl-Histone H3(K36) (D5A7) XP(R) Rabbit mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using Tri-Methyl-Histone H3 (Lys36) (D5A7) XP® Rabbit mAb (green) and COX IV (4D11-B3-E8) Mouse mAb #11967 (red).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HeLa cells using Tri-Methyl-Histone H3 (Lys36) (D5A7) XP® Rabbit mAb (blue) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.


Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either Tri-Methyl-Histone H3 (Lys36) (D5A7) XP® 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 γ-Actin Promoter Primers #5037, SimpleChIP® Human γ-Actin Intron 3 Primers #5047, SimpleChIP® Human GAPDH Promoter Primers #4471, and SimpleChIP® Human GAPDH Intron 2 Primers #4478. 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
 
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SignalStain is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

本製品は試験研究用です。

Tri-Methyl-Histone H3 (Lys36) (D5A7) XP® Rabbit mAb

Immune Cell Signaling Pathways

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