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#46599 Methyl-Histone H3 (Lys36) Antibody Sampler Kit

 
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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 (Lys36) (D5A7) XP® Rabbit mAb #4909 20 µl WB, IHC-P, IF-IC, F, ChIP, ChIP-seq H, M, R, Mk Hm, C, Dm, X, Z, B 17 Rabbit IgG
Di-Methyl-Histone H3 (Lys36) (C75H12) Rabbit mAb #2901 20 µl WB, IHC-P, IF-IC, F H, M, R, Mk 17 Rabbit IgG
Mono-Methyl-Histone H3 (Lys36) (D9J1D) Rabbit mAb #14111 20 µl WB, IP, IF-IC, F, ChIP, ChIP-seq H, M, R, Mk Hm, B 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

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

Antibody specificity was determined by Western blotting. HeLa and NIH/3T3 cell extracts were probed with Di-Methyl Histone H3 (Lys36) (C75H12) Rabbit mAb alone (Panel A) or Di-Methyl-Histone H3 (Lys36) (C75H12) Rabbit mAb pre-adsorbed with 1.5 μM of various competitor peptides (Panels B-I). As shown, only the di-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.


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 (Lys36) (D9J1D) Rabbit mAb.

Chromatin IP-seq

Chromatin IP-seq

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and Tri-Methyl-Histone H3 (Lys36) (D5A7) 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 ACTG1/γ-Actin, a known target gene of H3K36me3 (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 (Lys36) (D9J1D) 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 figures show binding across WDR70 gene. For additional ChIP-seq tracks, please download the product data sheet.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Di-Methyl-Histone H3 (Lys36) (C75H12) Rabbit mAb.

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.


IHC-P (paraffin)

IHC-P (paraffin)

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

Western Blotting

Western Blotting

Antibody specificity was determined by western blotting. HeLa and NIH/3T3 cell extracts were probed with Mono-Methyl-Histone H3 (Lys36) (D9J1D) Rabbit mAb alone (A) or Mono-Methyl-Histone H3 (Lys36) (D9J1D) Rabbit mAb

pre-adsorbed with 1.5 μM of various competitor peptides (B-I). As shown, only the mono-methyl-histone H3 (Lys36)

peptide competed away binding of the antibody (C).

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either Mono-Methyl-Histone H3 (Lys36) (D9J1D) 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 AFM Intron 1 Primers #5098, SimpleChIP® Human GAPDH Exon 1 Primers #5516, SimpleChIP® Human MyoD Exon 1 Primers #4490, and SimpleChIP® Human MYT-1 Exon 1 Primers #4493. 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 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 gastric carcinoma using Di-Methyl-Histone H3 (Lys36) (C75H12) Rabbit mAb in the presence of non-methyl peptide (left) or K36 di-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.


Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HeLa cells using Mono-Methyl-Histone H3 (Lys36) (D9J1D) 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 colon carcinoma using Tri-Methyl-Histone H3(K36) (D5A7) XP(R) Rabbit mAb.

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).


IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using Mono-Methyl-Histone H3 (Lys36) (D9J1D) Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HeLa cells using Di-Methyl-Histone H3 (Lys36) (C75H12) 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.

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.


IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using Di-Methyl-Histone H3 (Lys36) (C75H12) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).

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.


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.

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).

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 Lys36 is associated with transcriptionally active genes. Tri- and di-methyl-histone H3 Lys36 levels are high in the bodies of active genes, where these marks function to repress intragenic transcription initiation and regulate mRNA splicing. Mono-methyl-histone H3 Lys36 levels are high in the bodies of both active and inactive genes.

使用例
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関連製品
14111   Mono-Methyl-Histone H3 (Lys36) (D9J1D) Rabbit mAb
2901   Di-Methyl-Histone H3 (Lys36) (C75H12) Rabbit mAb
4499   Histone H3 (D1H2) XP® Rabbit mAb
4909   Tri-Methyl-Histone H3 (Lys36) (D5A7) XP® Rabbit mAb
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
9999   Nonfat Dry Milk

Illumina is a registered trademark of Illumina, Inc.
Tween is a registered trademark of ICI Americas, Inc.
SignalStain 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.
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 (Lys36) Antibody Sampler Kit

Metabolic Reprogramming in Disease

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