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Histone H3 [Asym-dimethyl Arg9] Antibody

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-44066

Novus Biologicals, part of Bio-Techne
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NBP2-44066

Key Product Details

Species Reactivity

Validated:

Human, Mouse, C. elegans

Applications

Chromatin Immunoprecipitation (ChIP), Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Product Summary for Histone H3 [Asym-dimethyl Arg9] Antibody

Immunogen

Histone H3 [Asym-dimethyl Arg9] Antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic dimethylated peptide surrounding Arginine 8 of human Histone H3.2. (Uniprot: Q71DI3)

Reactivity Notes

Predicted to react with many species including rat, chicken, Xenopus, Drosophila, and plant based on 100% sequence homology. Cross-reactivity with Histone H3 from other sources has not been determined

Modification

Asym-dimethyl Arg9

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Description

This antibody was affinity purified from monospecific antiserum by immunoaffinity chromatography

Store vial at -20C prior to opening. Aliquot contents and freeze at -20C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4C as an undiluted liquid. Dilute only prior to immediate use.

Scientific Data Images for Histone H3 [Asym-dimethyl Arg9] Antibody

Chromatin Immunoprecipitation: Histone H3 [Asym-dimethyl Arg9] Antibody [NBP2-44066] - Chromatin from one million formaldehyde cross-linked Hela cells was used with 2ug of Anti-Histone H3 R8me2a and 20ul of magnetic IgG beads per immunoprecipitation. A no antibody (No Ab) control was also used. Immunoprecipitated DNA was quantified using quantitative real-time PCR and SYBR green dye, then normalized to the non-precipitated input chromatin, which is equal to one.
Western Blot: Histone H3 [Asym-dimethyl Arg9] Antibody [NBP2-44066]

Western Blot: Histone H3 [Asym-dimethyl Arg9] Antibody [NBP2-44066]

Western Blot: Histone H3 [Asym-dimethyl Arg9] Antibody [NBP2-44066] - lane 1: HeLa Histone Prep Lysate. 2. NIH-3T3 Prep Lysate. Lane 3: C. elegans embryo lysate. Load: 30 Ug per lane. Observed molecular weight is ~16 kDa.
Immunocytochemistry/ Immunofluorescence: Histone H3 [Asym-dimethyl Arg9] Antibody [NBP2-44066]

Immunocytochemistry/ Immunofluorescence: Histone H3 [Asym-dimethyl Arg9] Antibody [NBP2-44066]

Immunocytochemistry/Immunofluorescence: Histone H3 [Asym-dimethyl Arg9] Antibody [NBP2-44066] - Tissue: HeLa cells. Fixation: 0.5% PFA. Antigen retrieval: Not required.

Applications for Histone H3 [Asym-dimethyl Arg9] Antibody

Application
Recommended Usage

Chromatin Immunoprecipitation (ChIP)

2-5ug/million cells

Immunocytochemistry/ Immunofluorescence

1:200

Immunohistochemistry

1:200

Western Blot

1:500
Application Notes
This product is tested for Western Blot, Immunocytochemistry, Immunofluorescence, Chromatin Immunoprecipitation, and Dot Blot. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately ~15.4 kDa corresponding to Histone H3 protein by Western Blotting in HeLa histone prep lysate or the appropriate cell lysate or extract. Epi-Plus(TM) antibody production in collaboration with Novus Biologicals.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 30% Glycerol

Preservative

0.01% Sodium Azide

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at -20C short term. Aliquot and store at -80C long term. Avoid freeze-thaw cycles.

Background: Histone H3

Chromatin is an arrangement of DNA and proteins which form chromosomes. Chromatin is formed from nucleosomes, which are comprised of a set of four histone proteins (H2A, H2B, H3, H4) wrapped with DNA. Chromatin is very dynamic, where post-translational modifications can regulate DNA to be copied, transcribed, or repaired.

Histones are nuclear proteins responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Changes in chromatin structure play a large role in the regulation of transcription. The chromatin fibers are compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures.

Common histone modifications include methylation of lysine and arginine, acetylation of lysine, phosphorylation of threonine and serine, and sumoylation, biotinylation, and ubiquitylation of lysine. Posttranslational modifications such as acetylation of core histones regulates gene expression, thus altering protein function and regulation (1). Histone H3 is primarily acetylated at lysines 9, 14, 18, and 23 and have a theoretical molecular weight of 15 kDa. Acetylation at lysine 9 and 14 appears to control histone deposition, chromatin assembly and active transcription. Methylation of arginine residues within histone H3 has also been linked to transcription regulation. Histone H3 has been linked to various types of cancer as a biomarker through the aberrant expression of histone deacetylase (HDAC) enzymes and changes to chromatins (2-4).

References

1. Zhang, Y. X., Akumuo, R. C., Espana, R. A., Yan, C. X., Gao, W. J., & Li, Y. C. (2018). The histone demethylase KDM6B in the medial prefrontal cortex epigenetically regulates cocaine reward memory. Neuropharmacology, 141, 113-125. doi:10.1016/j.neuropharm.2018.08.030

2. Nandakumar, V., Hansen, N., Glenn, H. L., Han, J. H., Helland, S., Hernandez, K, ...Meldrum, D. R. (2016). Vorinostat differentially alters 3D nuclear structure of cancer and non-cancerous esophageal cells. Sci Rep, 6, 30593. doi:10.1038/srep30593

3. Zhou, M., Li, Y., Lin, S., Chen, Y., Qian, Y., Zhao, Z., & Fan, H. (2019). H3K9me3, H3K36me3, and H4K20me3 Expression Correlates with Patient Outcome in Esophageal Squamous Cell Carcinoma as Epigenetic Markers. Dig Dis Sci, 64(8), 2147-2157. doi:10.1007/s10620-019-05529-2

4. Li, Y., Guo, D., Sun, R., Chen, P., Qian, Q., & Fan, H. (2019). Methylation Patterns of Lys9 and Lys27 on Histone H3 Correlate with Patient Outcome in Gastric Cancer. Dig Dis Sci, 64(2), 439-446. doi:10.1007/s10620-018-5341-8

Alternate Names

H3F3A, H3R9Me2a

Gene Symbol

H3C14

UniProt

Additional Histone H3 Products

Product Documents for Histone H3 [Asym-dimethyl Arg9] Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for Histone H3 [Asym-dimethyl Arg9] Antibody

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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