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#26482 Methyl-Histone H3 (Lys4) Antibody Sampler Kit

 
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希望納入価格 (円)
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2019年1月17日15時10分 現在
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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 (Lys4) (C42D8) Rabbit mAb #9751 20 µl WB, IHC-P, IF-IC, F, ChIP, ChIP-seq H, M, R, Mk, Dm, Sc X, Z 17 Rabbit IgG
Di-Methyl-Histone H3 (Lys4) (C64G9) Rabbit mAb #9725 20 µl WB, IP, IHC-P, IF-IC, F, ChIP, ChIP-seq H, M, R, Mk 17 Rabbit IgG
Mono-Methyl-Histone H3 (Lys4) (D1A9) XP® Rabbit mAb #5326 20 µl WB, IF-IC, F, ChIP, ChIP-seq H, M, R, Mk Dm 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, IHC-P=Immunohistochemistry (Paraffin), IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry, ChIP=Chromatin IP, ChIP-seq=Chromatin IP-seq, IP=Immunoprecipitation
Reactivity Key: H=Human, M=Mouse, R=Rat, Mk=Monkey, Dm=D. melanogaster, Sc=S. cerevisiae

貯法
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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 whole cell lysates from HeLa, NIH/3T3, C6 and COS cells using Di-Methyl-Histone H3 (Lys4) (C64G9) Rabbit mAb.

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

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Mono-Methyl-Histone H3 (Lys4) (D1A9) XP® Rabbit mAb.

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-seq

Chromatin IP-seq

Chromatin immunoprecipitations were performed with cross-linked chromatin from Hela cells and Mono-Methyl-Histone H3 (Lys4) (D1A9) XP® 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 ADAM9, a known target gene of H3K4me1 (see additional figure 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 Di-Methyl-Histone H3 (Lys4) (C64G9) Rabbit mAb, using SimpleChIP® 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 H3K4me2 (see additional figure containing ChIP-qPCR data). 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 Di-Methyl-Histone H3 (Lys4) (C64G9) Rabbit mAb.


Western Blotting

Western Blotting

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

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either Di-Methyl-Histone H3 (Lys4) (C64G9) 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.

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.


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 K562 cells and either Mono-Methyl-Histone H3 (Lys4) (D1A9) 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 ADAM9 Intron 11 Primers #73401, human ADAM18 intron 14 primers, human Trio inton 1 primers, and SimpleChIP® Human Trio Exon 57 Primers #90568. 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 HeLa cells using Mono-Methyl-Histone H3 (Lys4) (D1A9) XP® Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) #4414 was used as a secondary antibody.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human stomach carcinoma using Di-Methyl-Histone H3 (Lys4) (C64G9) Rabbit mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using Mono-Methyl-Histone H3 (Lys4) (D1A9) XP® Rabbit mAb (green) and MEK1/2 (L38C12) Mouse mAb #4694 (blue). Actin filaments were labeled with Dy-554 phalloidin (red).

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).


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 Di-Methyl-Histone H3 (Lys4) (C64G9) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).

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).


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.

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).

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 Lys4 is associated with transcriptional activation. Mono-methyl-histone H3 Lys4 levels are high at trancriptional enhancer elements, with lower levels of mono-methylation found at the promoters of active genes. Tri-methyl-histone H3 Lys4 levels are high at the promoters of active genes, in addition to bivalent, transcriptionally poised genes that also contain the repressive tri-methyl-histone H3 Lys27 modification. Di-methyl-histone H3 Lys4 levels are highest in the 5'-end of transcriptionally active genes.

使用例
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関連製品
4499   Histone H3 (D1H2) XP® Rabbit mAb
5326   Mono-Methyl-Histone H3 (Lys4) (D1A9) 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
7727   Biotinylated Protein Ladder Detection Pack
9725   Di-Methyl-Histone H3 (Lys4) (C64G9) Rabbit mAb
9751   Tri-Methyl-Histone H3 (Lys4) (C42D8) Rabbit mAb
9999   Nonfat Dry Milk

Illumina is a registered trademark of Illumina, Inc.
Tween is a registered trademark of ICI Americas, Inc.
XP 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.
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。

Methyl-Histone H3 (Lys4) Antibody Sampler Kit

Immune Cell Signaling Pathways

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