Histone H3 [Trimethyl Lys36] Antibody

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-59199

Novus Biologicals, part of Bio-Techne
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NBP2-59199
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Key Product Details

Species Reactivity

Validated:

Human, Drosophila, Zebrafish

Cited:

Human

Applications

Validated:

Chromatin Immunoprecipitation (ChIP), Chromatin Immunoprecipitation Sequencing, Dot Blot, ELISA, Western Blot

Cited:

Chemotaxis, Chromatin Immunoprecipitation (ChIP)

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Concentration

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

View all Histone H3 Antibodies »

Product Specifications

Immunogen

The exact sequence of the immunogen to this Histone H3 [Trimethyl Lys36] antibody is proprietary.

Modification

Trimethyl Lys36

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

15 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for Histone H3 [Trimethyl Lys36] Antibody

Western Blot: Histone H3 [Trimethyl Lys36] Antibody [NBP2-59199]

Western Blot: Histone H3 [Trimethyl Lys36] Antibody [NBP2-59199]

Western Blot: Histone H3 [Trimethyl Lys36] Antibody [NBP2-59199] - Histone extracts (15 ug) from HeLa cells were analysed by Western blot using the antibody directed against H3K36me3 diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. Observed molecular weight is ~17 kDa.
Chromatin Immunoprecipitation: Histone H3 [Trimethyl Lys36] Antibody [NBP2-59199] - ChIP assays were performed using HeLa cells, the antibody against H3K36me3 and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 1 million cells. A titration consisting of 1, 2, 5 and 10 ug of antibody per ChIP experiment was analyzed. IgG (2 ug/IP) was used as a negative IP control. QPCR was performed with primers for the promoter and coding region of the active GAPDH, for a region located 1 kb upstream of the GAPDH promoter and for the Sat2 satellite repeat. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
ELISA: Histone H3 [Trimethyl Lys36] Antibody [NBP2-59199]

ELISA: Histone H3 [Trimethyl Lys36] Antibody [NBP2-59199]

ELISA: Histone H3 [Trimethyl Lys36] Antibody [NBP2-59199] - To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody directed against H3K36me3 and the crude serum. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the purified antibody was estimated to be 1:19,300.
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Applications for Histone H3 [Trimethyl Lys36] Antibody

Application
Recommended Usage

Chromatin Immunoprecipitation (ChIP)

1-2 ug

Dot Blot

1:20000

ELISA

1:500

Western Blot

1:1000
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Peptide affinity purified

Formulation

PBS

Preservative

0.05% Sodium Azide and 0.05% ProClin 300

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. 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, H3K36Me3

Gene Symbol

H3C14

Additional Histone H3 Products

Product Documents for Histone H3 [Trimethyl Lys36] Antibody

Product Datasheets

Certificate of Analysis

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Product Specific Notices for Histone H3 [Trimethyl Lys36] 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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