Histone H3 [Dimethyl Lys4] Antibody (JE00-98)

Name: Histone H3 [Dimethyl Lys4] Antibody (JE00-98)
Brand: Novus Biologicals
SKU: NBP3-32431
Price: 439 USD
Availability: InStock

Novus Biologicals | Catalog # NBP3-32431

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Product Documents

Recombinant Monoclonal Antibody

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Product Specifications
Scientific Data
Applications
Preparation & Storage
Background
Product Documents
Specific Notices
Research Areas
Related Blogs

Key Product Details

Species Reactivity

Human, Mouse, Rat, Monkey

Applications

Western Blot, Immunocytochemistry/ Immunofluorescence, Chromatin Immunoprecipitation, Dot Blot

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # JE00-98 expressed in HEK293

View all Histone H3 Primary Antibodies »

Product Specifications

Immunogen

Synthetic peptide corresponding to the amino terminus of histone H3 in which Lys4 is di-methylated. (Uniprot: P68431)

Modification

Dimethyl Lys 4

Localization

Nucleus, Chromosome.

Clonality

Monoclonal

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 [Dimethyl Lys4] Antibody (JE00-98)

Chromatin Immunoprecipitation: Histone H3 [Dimethyl Lys4] Antibody (JE00-98) [NBP3-32431]

Chromatin Immunoprecipitation: Histone H3 [Dimethyl Lys4] Antibody (JE00-98) [NBP3-32431] - Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells and either Histone H3 (NBP3-32431) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

Western Blot: Histone H3 [Dimethyl Lys4] Antibody (JE00-98) [NBP3-32431]

Western Blot: Histone H3 [Dimethyl Lys4] Antibody (JE00-98) [NBP3-32431] - Western blot analysis of Histone H3 on different lysates with Rabbit anti-Histone H3 antibody (NBP3-32431) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: MCF7 cell lysate
Lane 3: HCT 116 cell lysate
Lane 4: COS-1 cell lysate
Lane 5: NIH/3T3 cell lysate
Lane 6: C6 cell lysate

Lysates/proteins at 10 ug/Lane.

Predicted band size: 15 kDa
Observed band size: 15 kDa

Exposure time: 14 seconds; ECL;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (NBP3-32431) at 1/1,000 dilution was used in 5% NFDM/TBST at 4 overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.

Dot Blot: Histone H3 [Dimethyl Lys4] Antibody (JE00-98) [NBP3-32431]

Dot Blot: Histone H3 [Dimethyl Lys4] Antibody (JE00-98) [NBP3-32431] - Dot blot analysis of Histone H3 on different proteins with Rabbit anti-Histone H3 antibody (NBP3-32431) at 1/1,000 dilution. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution for 1 hour at room temperature.

Lane 1: Unmodified Histone H3 (negative)
Lane 2: Mono-Methyl-Histone H3 (Lys4) (negative)
Lane 3: Di-Methyl-Histone H3 (Lys4) (positive)
Lane 4: Tri-Methyl-Histone H3 (Lys4) (negative)
Lane 5: Acetyl-Histone H3 (Lys4) (negative)
Lane 6: Acetyl-Histone H3 (Lys9) (negative)

Proteins loading: 100ng, 25ng, 5ng, 1ng;

Blocking and dilution buffer: 5% NFDM/TBST;

Exposure time: 15 seconds; ECL.

Immunohistochemistry: Histone H3 [Dimethyl Lys4] Antibody (JE00-98) [NBP3-32431]

Immunohistochemistry: Histone H3 [Dimethyl Lys4] Antibody (JE00-98) [NBP3-32431] - Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-Histone H3 antibody (NBP3-32431) at 1/10,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (NBP3-32431) at 1/10,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunocytochemistry/ Immunofluorescence: Histone H3 [Dimethyl Lys4] Antibody (JE00-98) [NBP3-32431]

Immunocytochemistry/ Immunofluorescence: Histone H3 [Dimethyl Lys4] Antibody (JE00-98) [NBP3-32431] - Immunocytochemistry analysis of HeLa cells labeling Histone H3 with Rabbit anti-Histone H3 antibody (NBP3-32431) at 1/500 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Histone H3 antibody (NBP3-32431) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.

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Applications for Histone H3 [Dimethyl Lys4] Antibody (JE00-98)

Application

Recommended Usage

Chromatin Immunoprecipitation

Use 0.5-2 ug for 25 ug of chromatin

Dot Blot

1:1000

Immunocytochemistry/ Immunofluorescence

1:500-1:2000

Western Blot

1:1000

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

PBS (pH7.4), 0.1% BSA and 40% Glycerol

Preservative

0.05% Sodium Azide

Concentration

1 mg/ml

Shipping

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

Stability & Storage

Store at 4C short term. Aliquot and store at -20C 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

Gene Symbol

H3C14

Additional Histone H3 Products

Product Documents for Histone H3 [Dimethyl Lys4] Antibody (JE00-98)

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Certificate of Analysis

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Product Specific Notices for Histone H3 [Dimethyl Lys4] Antibody (JE00-98)

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.

Related Research Areas

Epigenetics

Interferons

Notch Pathway

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