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#12799 Acetyl-Histone H2B (Lys5) (D5H1S) XP® Rabbit mAb

 
CSTコード 包装
希望納入価格 (円)
国内在庫 i
2019年11月15日15時30分 現在
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#12799S100 μL72,000
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#12799T20 μL39,000
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HISTONE H2B 製品一覧 | #12799 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14Rabbit 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、ゼブラフィッシュ、ウシ、ウマ
12799 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)免疫沈降 (1:100)免疫組織染色 (パラフィン) (1:1000)免疫蛍光細胞染色 (IF-IC) (1:400)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Lys5 がアセチル化されたHistone H2B タンパク質を検出します。他のアセチル化Histone タンパク質とは交差しません。
使用抗原
ヒトのHistone H2B タンパク質のアセチル化されたLys5 周辺領域 (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12799 の推奨プロトコールで実験した結果です。

ELISA-Peptide

ELISA-Peptide

Acetyl-Histone H2B (Lys5) (D5H1S) XP® Rabbit mAb specificity was determined by peptide ELISA. The graph depicts the binding of the antibody to pre-coated acetyl-histone H2B (Lys5) peptide in the presence of increasing concentrations of various competitor peptides. As shown, only the acetyl-histone H2B (Lys5) peptide competed away binding of the antibody.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa and C6 cells, untreated (-) or treated with Trichostatin A (TSA) #9950 (1 μM, 18hr; +), using Acetyl-Histone H2B (Lys5) (D5H1S) XP® Rabbit mAb (upper) or Histone H2B (D2H6) Rabbit mAb #12364 (lower).

IP

IP

Immunoprecipitation of acetylated histone H2B from HeLa cell extracts treated with Trichostatin A (TSA) #9950 (1 μM, 18hr) using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Acetyl-Histone H2B (D5H1S) XP® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Acetyl-Histone H2B (D5H1S) XP® Rabbit mAb.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Acetyl-Histone H2B (Lys5) (D5H1S) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Acetyl-Histone H2B (Lys5) (D5H1S) XP® Rabbit mAb in the presence of non-acetyl peptide (left) or Lys5 acetyl peptide (right).

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (right) or treated with Trichostatin A (TSA) #9950 (left), using Acetyl-Histone H2B (Lys5) (D5H1S) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).


Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either Acetyl-Histone H2B (Lys5) (D5H1S) 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 GAPDH Exon 1 Primers #5516, SimpleChIP® Human RPL30 Exon 3 Primers #7014, 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 (1,2). The p300/CBP histone acetyltransferases acetylate multiple lysine residues in the amino terminal tail of histone H2B (Lys5, 12, 15, and 20) at gene promoters during transcriptional activation (1-3). Hyper-acetylation of the histone tails neutralizes the positive charge of these domains and is believed to weaken histone-DNA and nucleosome-nucleosome interactions, thereby destabilizing chromatin structure and increasing the access of DNA to various DNA-binding proteins (4,5). In addition, acetylation of specific lysine residues creates docking sites that facilitate recruitment of many transcription and chromatin regulatory proteins that contain a bromodomain, which binds to acetylated lysine residues (6). Histone H2B is mono-ubiquitinated at Lys120 during transcriptional activation by the RAD6 E2 protein in conjunction with the BRE1A/BRE1B E3 ligase (also known as RNF20/RNF40) (7). Mono-ubiquitinated histone H2B Lys120 is associated with the transcribed region of active genes and stimulates transcriptional elongation by facilitating FACT-dependent chromatin remodeling (7-9). In addition, it is essential for subsequent methylation of histone H3 Lys4 and Lys79, two additional histone modifications that regulate transcriptional initiation and elongation (10). In response to metabolic stress, AMPK is recruited to responsive genes and phosphorylates histone H2B at Lys36, both at promoters and in transcribed regions of genes, and may regulate transcriptional elongation (11). In response to multiple apoptotic stimuli, histone H2B is phosphorylated at Ser14 by the Mst1 kinase (12). Upon induction of apoptosis, Mst1 is cleaved and activated by caspase-3, leading to global phosphorylation of histone H2B during chromatin condensation. Interestingly, histone H2B is rapidly phosphorylated at irradiation-induced DNA damage foci in mouse embryonic fibroblasts (13). In this case, phosphorylation at Ser14 is rapid, depends on prior phosphorylation of H2AX Ser139, and occurs in the absence of apoptosis, suggesting that Ser14 phosphorylation may have distinct roles in DNA-damage repair and apoptosis.

使用例
 
関連製品
12364   Histone H2B (D2H6) Rabbit mAb
12630   SignalFire™ Plus ECL Reagent
12757   SignalFire™ Elite ECL Reagent
2571   Acetyl-Histone H2B (Lys20) Antibody
4486   SimpleChIP® Human α Satellite Repeat Primers
4490   SimpleChIP® Human MyoD1 Exon 1 Primers
5516   SimpleChIP® Human GAPDH Exon 1 Primers
6883   SignalFire™ ECL Reagent
7014   SimpleChIP® Human RPL30 Exon 3 Primers
7017   6-Tube Magnetic Separation Rack
7074   Anti-rabbit IgG, HRP-linked Antibody
7727   Biotinylated Protein Ladder Detection Pack
8059   SignalStain® DAB Substrate Kit
8112   SignalStain® Antibody Diluent
8114   SignalStain® Boost IHC Detection Reagent (HRP, Rabbit)
8135   Histone H2B (V119) Antibody
9002   SimpleChIP® Enzymatic Chromatin IP Kit (Agarose Beads)
9003   SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads)
9004   SimpleChIP® Plus Enzymatic Chromatin IP Kit (Agarose Beads)
9005   SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads)
9006   ChIP-Grade Protein G Magnetic Beads
9007   ChIP-Grade Protein G Agarose Beads
9701   Phospho-Histone H3 (Ser10) Antibody
9706   Phospho-Histone H3 (Ser10) (6G3) Mouse mAb
9711   Acetyl- and Phospho-Histone H3 (Lys9/Ser10) Antibody
9715   Histone H3 Antibody
9997   Tris Buffered Saline with Tween® 20 (TBST-10X)

DRAQ5 is a registered trademark of Biostatus Limited.
XP is a registered trademark of Cell Signaling Technology, Inc.
SimpleChIP 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。

Acetyl-Histone H2B (Lys5) (D5H1S) XP® Rabbit mAb

Metabolic Reprogramming in Disease

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