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#45139 Methyl-Histone H3 (Lys79) Antibody Sampler Kit

 
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2019年2月22日15時50分 現在
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#45139T1 Kit75,000
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キット内容 容量 用途 種交差性 ホモロジー† 検出分子量 アイソタイプ
Tri-Methyl-Histone H3 (Lys79) (E8B3M) Rabbit mAb #74073 20 µl WB, ChIP, ChIP-seq H, M, R, Mk 17 Rabbit IgG
Di-Methyl-Histone H3 (Lys79) (D15E8) XP® Rabbit mAb #5427 20 µl WB, ChIP H, M, R, Mk 17 Rabbit IgG
Mono-Methyl-Histone H3 (Lys79) (D5X1S) Rabbit mAb #12522 20 µl WB, ChIP, ChIP-seq H, M, R, Mk 17 Rabbit IgG
Histone H3 (D1H2) XP® Rabbit mAb #4499 20 µl WB, IHC-P, IF-IC, F H, M, R, Mk Hm, C, Dm, X, Z, B 17 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl WB Goat

†Species predicted to react based on 100% sequence homology.

Applications Key: W=Western Blotting, ChIP=Chromatin IP, ChIP-seq=Chromatin IP-seq, IHC-P=Immunohistochemistry (Paraffin), IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry
Reactivity Key: H=Human, M=Mouse, R=Rat, Mk=Monkey

特異性・感度
Each antibody in the Methyl-Histone H3 (Lys79) Antibody Sampler Kit detects endogenous levels of its target protein. Tri-Methyl-Histone H3 (Lys79) (E8B3M) Rabbit mAb recognizes endogenous levels of histone H3 protein when tri-methylated at Lys79. This antibody shows some cross-reactivity to histone H3 that is di-methylated on Lys79, but does not cross-react with non-methylated or mono-methylated histone H3 Lys79. Di-Methyl-Histone H3 (Lys79) (D15E8) XP® Rabbit mAb detects endogenous levels of histone H3 only when di-methylated on Lys79. Mono-Methyl-Histone H3 (Lys79) (D5X1S) Rabbit mAb recognizes endogenous levels of histone H3 protein only when mono-methylated at Lys79. These methyl-Lys79 antibodies do not cross-react with histone H3 methylated at Lys4, Lys9, Lys27, or Lys36. Histone H3 (D1H2) XP® Rabbit mAb detects endogenous levels of total Histone H3 protein, including isoforms H3.1, H3.2, H3.3, and the variant histone CENP-A. This antibody does not cross-react with other core histones.In addition, the antibody does not cross-react with methylated histone H3 Lys4, Lys9, Lys27, Lys36 or methylated histone H4 Lys20.
使用抗原
Monoclonal methyl-histone H3 Lys79 antibodies are produced by immunizing rabbits with synthetic peptides corresponding to the amino terminus of histone H3 in which Lys79 is mono-, di-, or tri-methylated. The control histone H3 monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the carboxy terminus of the human histone H3 protein.

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貯法
-20℃
社内データ

Western Blotting

Western Blotting

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Histone H3 (D1H2) XP® Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa and NIH/3T3 cell lines using Di-Methyl-Histone H3 (Lys79) (D15E8) XP® Rabbit mAb.


Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Mono-Methyl-Histone H3 (Lys79) (D5X1S) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Tri-Methyl-Histone H3 (Lys79) (E8B3M) Rabbit mAb.

Chromatin IP-seq

Chromatin IP-seq

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and Tri-Methyl-Histone H3 (Lys79) (E8B3M) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across GAPDH, a known target gene of H3K79me3 (see additional figures containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.


Chromatin IP-seq

Chromatin IP-seq

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and Mono-Methyl-Histone H3 (Lys79) (D5X1S) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across RAB5C gene. For additional ChIP-seq tracks, please download the product data sheet.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Histone H3 (D1H2) XP® Rabbit mAb.

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either Di-Methyl-Histone H3 (Lys79) (D15E8) 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.


Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin fromHeLa cells and either Mono-Methyl-Histone H3 (Lys79) (D5X1S) 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 Promoter Primers #4471, SimpleChIP® Human GAPDH Intron 2 Primers #4478, human PABPC1 promoter primers, and human PABPC1 intron 4 primers. 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.

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either Tri-Methyl-Histone H3 (Lys79) (E8B3M) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human RPL30 Exon 3 Primers #7014, SimpleChIP® Human GAPDH Promoter Primers #4471, and SimpleChIP® Human MyoD1 Exon 1 Primers #4490. 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.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of human peripheral blood lymphocytes using Histone H3 (D1H2) XP® Rabbit mAb (blue) compared to 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.


IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using Histone H3 (D1H2) XP® Rabbit mAb (green) and β-Tubulin (9F3) Rabbit mAb (Alexa Fluor® 555 Conjugate) #2116 (red).

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).

Methylation of histone H3 Lys79 is mediated by the DOT1L histone methyltransferase, is associated with transcriptionally active genes, and plays a role in the regulation of DNA damage response, cell cycle, and embryonic stem cell development.

使用例
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関連製品
12522   Mono-Methyl-Histone H3 (Lys79) (D5X1S) Rabbit mAb
4499   Histone H3 (D1H2) XP® Rabbit mAb
5427   Di-Methyl-Histone H3 (Lys79) (D15E8) XP® Rabbit mAb
7003   20X LumiGLO® Reagent and 20X Peroxide
7071   Phototope-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
7074   Anti-rabbit IgG, HRP-linked Antibody
74073   Tri-Methyl-Histone H3 (Lys79) (E8B3M) Rabbit mAb
7727   Biotinylated Protein Ladder Detection Pack
9999   Nonfat Dry Milk

Illumina is a registered trademark of Illumina, Inc.
Tween is a registered trademark of ICI Americas, Inc.
CST is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
LumiGLO is a registered trademark of Kirkegaard & Perry Laboratories.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

本製品は試験研究用です。

Methyl-Histone H3 (Lys79) Antibody Sampler Kit

Immune Cell Signaling Pathways

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