修饰抗体 Products
组蛋白赖氨酸二甲基化 (di-methylation) 修饰抗体
杭州景杰生物科技有限公司
Anti-di-methyl-Histone H3 (Lys9) rabbit pAb

Cat#:

PTM-615

Ab Type:

P

Applications:

WB, Dot, ELISA, IHC, ICC, IP, ChIP

Species:

Human, Mouse, Rat, Rice, Cerastium

Price: (100 μl)

2500

Price: (20 μl)

750.00


Product Description

The rabbit-derived antibody is purified with protein A-conjugated agarose followed by di-methylated histone H3 (Lys9) peptide affinity chromatography. It specifically recognizes histone H3 with di-methylation at Lys9.  


Source/Purification

This product is produced by immunizing rabbits with a synthetic di-methyl peptide corresponding to residues surrounding Lys9 of human histone H3. Antibodies are purified by protein A-conjugated agarose followed by di-methylated histone H3 (Lys9) peptide affinity chromatography.  


Specificity

H3

Recommended Applications

ELISA,WB, IHC, IP, ChIP; Not tested in other applications   

*WB=Western blotting IHC=immunohistochemistry    ICC=Immunocytochemistry

  ChIP=Chromatin Immunoprecipitation   IP=Immunoprecipitation

Recommended antibody dilution

WB: 1:2000    IHC: 1:500    ICC: 1:200    IP: 1:25~1:100

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.  


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Anti-di-methyl-Histone H3 (Lys9) rabbit pAb

D. Western blotting analysis on 30 µg of crude proteins from  Rice, Cerastium arvense  histone lysates, N2a and Spleen whole cell lysates  using di-methyl-histone H3 (Lys9) rabbit pAb (1:400).

Anti-di-methyl-Histone H3 (Lys9) rabbit pAb

D. Immunocytochemistry analysis of Hela (Human cervix adenocarcinoma cell line), labeling H3K9me2 with PTM-615 at 1/200 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Nuclei were stained with DAPI (blue).

Anti-di-methyl-Histone H3 (Lys9) rabbit pAb

E. Immunohistochemical analysis of paraffin-embedded mouse colon tissue, labeling H3K9me2 with PTM-615 at 1/500 dilution. The PTM-615 was pretreated with un-methyl-Histone H3 peptide (Panel A) and di-methyl-Histone H3 (Lys9) (Panel B) site-specific peptide for 2 hours.

Nuclear staining on epithelial cells in Panel A, no staining in Panel B. Counter stained with hematoxylin.

Anti-di-methyl-Histone H3 (Lys9) rabbit pAb

F. Chromatin prepared from 1 x 105 HeLa cells was subjected to chromatin immunoprecipitation (ChIP) using either serial dilutions of  Di-methyl-Histone H3 (Lys9) 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.


Recommended Applications

ELISA,WB, IHC, IP, ChIP; Not tested in other applications   

*WB=Western blotting IHC=immunohistochemistry    ICC=Immunocytochemistry

  ChIP=Chromatin Immunoprecipitation   IP=Immunoprecipitation

Recommended antibody dilution

WB: 1:2000    IHC: 1:500    ICC: 1:200    IP: 1:25~1:100


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