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Histone H3K9me3 (H3K9 Trimethyl) Polyclonal Antibody

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Citations (8) | (1) | Write a Review
Western blot analysis of extracts of HeLa cell line and H3 protein expressed in E.coli., using H3K9me3 Polyclonal Antibody.
Dot-blot analysis of all sorts of methylation peptides using H3K9me3 Polyclonal Antibody.
Immunohistochemistry of paraffin-embedded rat lung tissue using H3K9me3 Polyclonal Antibody at dilution of 1:200 (x400 lens).
Immunohistochemistry of paraffin-embedded human kidney tissue using H3K9me3 Polyclonal Antibody at dilution of 1:200 (x400 lens).
Immunohistochemistry of paraffin-embedded human lung cancer tissue using H3K9me3 antibody at dilution of 1:200 (x400 lens).
Immunohistochemistry of paraffin-embedded human liver cancer tissue using H3K9me3 Polyclonal Antibody at dilution of 1:200 (x400 lens).
Immunofluorescence analysis of 293T cell using H3K9me3 Polyclonal Antibody. Blue: DAPI for nuclear staining.
Chromatin immunoprecipitation analysis extracts of 293 cell line, using Histone H3K9 Trimethyl Polyclonal Antibody and rabbit IgG. The amount of immunoprecipitated DNA was checked by quantitative PCR. Histogram was constructed by the ratios of the immunoprecipitated DNA to the input.
Application: ChIP, ELISA, IF, IHC, IP, WB
Areas of Research: Histone Methylation & Demethylation
Clonality: Polyclonal
Conjugate: Unconjugated
Host: Rabbit
Isotype: IgG
Purification: Affinity Purified
Reactivity: Broad Range, Human, Mouse, Rat
100% Guarantee: 6 months
Catalog No.SizePriceQty
A-4036-02525 µg $143.00 
A-4036-05050 µg $277.00 
Availability: Usually Ships In 1-2 Days 
Product Overview

Background 
Modulation of chromatin structure plays an important role in the regulation of transcription in eukaryotes. The nucleosome, made up of DNA wound around eight core histone proteins (two each of H2A, H2B, H3, and H4), is the primary building block of chromatin (1). The amino-terminal tails of core histones undergo various post-translational modifications, including acetylation, phosphorylation, methylation, and ubiquitination (2-5). These modifications occur in response to various stimuli and have a direct effect on the accessibility of chromatin to transcription factors and, therefore, gene expression (6). In most species, histone H2B is primarily acetylated at Lys5, 12, 15, and 20 (4,7). Histone H3 is primarily acetylated at Lys9, 14, 18, 23, 27, and 56. Acetylation of H3 at Lys9 appears to have a dominant role in histone deposition and chromatin assembly in some organisms (2,3). Phosphorylation at Ser10, Ser28, and Thr11 of histone H3 is tightly correlated with chromosome condensation during both mitosis and meiosis (8-10). Phosphorylation at Thr3 of histone H3 is highly conserved among many species and is catalyzed by the kinase haspin. Immunostaining with phospho-specific antibodies in mammalian cells reveals mitotic phosphorylation at Thr3 of H3 in prophase and its dephosphorylation during anaphase (11).

Concentration
1 mg/ml

Description
Rabbit polyclonal antibody raised against a synthetic peptide corresponding to the trimethylated histone H3 at K9, ChIP-grade

Purification
Affinity purification

Specificity
Broad Range, Mouse, Rat, Human

Isotype
IgG

Immunogen
A synthetic methylated peptide corresponding to residues surrounding K9 of human histone H3

Formulation 
Buffer: PBS with 0.02% sodium azide, 50% glycerol, pH7.3

Storage
Store at -20°C (regular) or -80°C (long term). Avoid freeze / thaw cycles.

Alternative Names
H3K9me3 antibody; H3K9m3 antibody; HIST1H3J; H3/j; H3FJ; Histone H3.1; Histone H3/a; Histone H3/b; Histone H3/c; Histone H3/d; Histone H3/f; Histone H3/h; Histone H3/I; Histone H3/j; Histone H3/k; Histone H3/l; HIST3H3 

Application 
WB: 1:500 - 1:2000
IHC: 1:50 - 1:200
IF: 1:50 - 1:200
IP: 1:50 - 1:200
ChIP: 1:50 - 1:200

User Guide & MSDS

[User Guide]*
*Always use the actual User Guide that shipped with your product. Is the above file locked? You can also request user guides by emailing info@epigentek.com along with your contact information and institution name.

Product Citations

Jiajia Xu et. al. (September 2016). Expression of SIRT1, H3K9me3, H3K9Ac and E-cadherin and its correlations with clinicopathological characteristics in gastric cancer patients IJCEP. 9(9)

Timoshevskiy VA et. al. (June 2016). Cellular and Molecular Features of Developmentally Programmed Genome Rearrangement in a Vertebrate (Sea Lamprey: Petromyzon marinus). PLoS Genet. 12(6):e1006103.

Trapphoff T et. al. (November 2015). Postovulatory aging affects dynamics of mRNA, expression and localization of maternal effect proteins, spindle integrity and pericentromeric proteins in mouse oocytes. Hum Reprod.

Tyler CR et. al. (July 2015). Developmental exposure to 50 parts-per-billion arsenic influences histone modifications and associated epigenetic machinery in a region- and sex-specific manner in the adult mouse brain. Toxicol Appl Pharmacol.

Wiegreffe C et. al. (July 2015). Bcl11a (Ctip1) Controls Migration of Cortical Projection Neurons through Regulation of Sema3c. Neuron. 87(2):311-25.

Kumar P et. al. (March 2014). Histone deacetylase inhibitors modulate the transcriptional regulation of guanylyl cyclase/natriuretic peptide receptor-a gene: interactive roles of modified histones, histone acetyltransferase, p300, AND Sp1. J Biol Chem. 289(10):6991-7002.

Sui WG et. al. (January 2014). ChIP-seq analysis of histone H3K9 trimethylation in peripheral blood mononuclear cells of membranous nephropathy patients. Braz J Med Biol Res. 47(1):42-9.

Trapphoff T et. al. (December 2013). Chronic exposure to a low concentration of bisphenol A during follicle culture affects the epigenetic status of germinal vesicles and metaphase II oocytes. Fertil Steril. 100(6):1758-67.e1.

Customer Reviews

Rating By J*************@ouhsc.edu Verified Purchase Reviewed on: Monday 13 March, 2017
Application Description
This antibody worked great for our experiment. The background was low, and the staining was specific to the nucleus.

Procedural Details
Histone H3K9me3 (H3K9 trimethyl) polyclonal antibody was used as a negative control for a co-localization experiment using immunofluorescence, as our protein of interest does not bind H3K9me3. Pre-B cells were attached to coverslips, fixed in 4% paraformaldehyde, and permeabilized using Triton X-100. Cells were then incubated with anti-Histone H3K9me3 1:100 for one hour, followed by secondary antibody staining with Cy3 anti-rabbit 1:250 for 45 minutes. DAPI was used as nuclear stain.

Other Thoughts
Pros:
Availability of multiple sizes/volumes of the product, relatively cheap compared to other companies, arrived fast, tested using multiple applications, very little background staining.

Cons:
None with the antibody in particular, we have little to no experience with Epigentek antibodies so we were hesitant when purchasing (we weren’t sure how good the antibody would be).


Immunofluorescence staining with the H3K9me3 polyclonal antibody.
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