修饰抗体 Products
组蛋白赖氨酸三甲基化 (tri-methylation) 修饰抗体
杭州景杰生物科技有限公司
Anti-tri-methyl-Histone H3 (Lys4) rabbit pAb

Cat#:

PTM-613

Ab Type:

P

Applications:

WB, Dot, IHC, ICC, ChIP

Species:

Human, Mouse, Rat

Price: (100 μl)

2500

Price: (20 μl)

750.00


Product Description

The rabbit-derived antibody is purified with protein A-conjugated agarose followed by tri-methylated histone H3 (Lys4) peptide affinity chromatography. It specifically recognizes histone H3 with tri-methylation at Lys4.  


Source/Purification

This product is produced by immunizing rabbits with a synthetic tri-methyl peptide corresponding to residues surrounding Lys4 of human histone H3. Antibodies are purified by protein A-conjugated agarose followed by tri-methylated histone H3 (Lys4) peptide affinitychromatography.  


Specificity

H3

Recommended Applications

ELISA,WB,IHC, ICC; Not tested in other applications 

*WB=Western blotting; IHC=Immunohistochemistry; ICC=Immunocytochemistry

  ChIP=Chromatin Immunoprecipitation


Recommended antibody dilution

WB: 1:2000, IHC:1:200, ICC:1:200

Important

For Western blotting, incubate  membrane with diluted antibody in 5% nonfat milk, 1 x TBS, 0.1% Tween-20 for  two hours at room temperature with gentle shaking.  

Use at an assay dependent concentration. Optimal  dilutions/concentrations should be determined by the end user.  




                            


              Anti-tri-methyl-Histone H3 (Lys4) rabbit pAb

Figure A. Dot  blotting analysis on indicated amount of tri-methylated H3 peptide at Lys4  (lane 1), mono-methylated H3 peptide at Lys4 (lane 2), di-methylated H3 peptide  at Lys4 (lane 3), mono-methylated H3 peptide at Lys9 (lane 4), di-methylated H3  peptide at Lys9 (lane 5), tri-methylated H3 peptide at Lys9 (lane 6),  mono-methylated H3 peptide at Lys14 (lane 7), di-methylated H3 peptide at Lys14  (lane 8), tri-methylated H3 peptide at Lys14 (lane 9) and unmodified H3 peptide  at Lys4 (lane 10) using tri-methyl-histone H3 (Lys4) rabbit pAb.

Anti-tri-methyl-Histone H3 (Lys4) rabbit pAb

Figure B. Western blotting analysis on 30 μg of crude proteins  from HeLa, C2C12 and COS-7 whole cell lysates using tri-methyl-histone H3 (Lys4)  rabbit pAb (1:2000).  

Anti-tri-methyl-Histone H3 (Lys4) rabbit pAb

Figure C. Immunohistochemistry analysis of paraffin-embedded mouse colon tissue, labeling H3K4me3 with PTM-613 at 1/500 dilution.

Nuclear staining on epithelial cells of mouse colon was observed. Counter stained with hematoxylin


Anti-tri-methyl-Histone H3 (Lys4) rabbit pAb

Figure D. Immunocytochemistry  analysis of 4% paraformaldehyde-fixed HeLa (human cervix adenocarcinoma epithelial cell) cells labeling tri-methyl-Histone H3 (Lys4) with PTM-613 at 1/200 dilution. Immunofluorescent image showed nuclear staining on HeLa cell line (Panel A). DAPI (blue) was used as the nuclear counterstain (Panel B).

Anti-tri-methyl-Histone H3 (Lys4) rabbit pAbChromatin prepared from 1 x 105 HeLa cells was subjected to chromatin immunoprecipitation (ChIP) using either serial dilutions of Tri-methyl-Histone H3 (Lys4) rabbit pAb or 1 μg of normal rabbit IgG as a negative control. Real time quantitative PCR was performed on immunoprecipitated DNA using primers specific for the human RPL30 Exon 3 and MyoD1 Exon 1. Data are presented as enrichment of each sample relative to total amount of input chromatin at each amplicon. 



Scientific Description

The ε-amino lysine acetylation of proteins is an important reversible modification controlling protein activity. The amino-terminal tails of core histones undergo lysine methylation in multiple sites, termed as “histone code” or “epigenetic code”. Lysine methylation in core histones is a major determinant for the formation of active and inactive regions of the genome and therefore plays vital roles in multiple cellular events. In most species, 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. Methylation in histones modulated by specific histone methylases (HMTs) and histone demethylases (HDMs) is impaired in the pathologies of cancer and other diseases and therefore, enzymes regulating histone lysine methylation have become promising targets for anti-cancer drugs.  


Storage & Stability

Store product at -20oC. Avoid repeated freeze/thaw. Antibody is supplied in PBS with 50% glycerol and 0.01% sodium azide.Stable for 12 months from date of receipt.


**Research purposes only.Not intended to be used for therapeutic or diagnostic purposes in humans or animals.



ELISA,WB,IHC; Not tested in other applications 

*WB=Western blotting  IHC=Immunohistochemistry  ICC=Immunocytochemistry

  ChIP=Chromatin Immunoprecipitation

WB: 1:2000    IHC: 1:200    ICC: 1:200



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