Key Product Details

Species Reactivity

Validated:

Human, Mouse, C. elegans

Cited:

Nematode - Caenorhabditis elegans

Predicted:

Chicken (100%), Drosophila (100%), Plant (100%), Rat (100%), Xenopus (100%). Backed by our 100% Guarantee.

Applications

Validated:

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

Cited:

IF/IHC, Immunocytochemistry/ Immunofluorescence

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free

Concentration

0.70 mg/ml

View all Histone H3 Antibodies »

Product Specifications

Immunogen

This Histone H3 [Dimethyl Lys4] antibody was raised against synthetic dimethylated peptide surrounding Lysine 4 of human Histone H3.2 [Swiss Prot Q71DI3].

Reactivity Notes

Predicted to react with many species including rat, chicken, Xenopus, Drosophila, and plant based on 100% sequence homology.

Modification

Dimethyl Lys4

Localization

Nucleus. Chromosome.

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 [Dimethyl Lys4] Antibody - BSA Free

Dot Blot: Histone H3 [Dimethyl Lys4] Antibody [NB21-1022] - Dot blot analysis of H3 K4me2 using the peptides stated above in 25, 10, 3 and 1 picomoles of peptide.
Chromatin Immunoprecipitation: Histone H3 [Dimethyl Lys4] Antibody [NB21-1022] - Chromatin from one million formaldehyde cross-linked Hela cells was used with 2ug of NB21-1022 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 [Dimethyl Lys4] AntibodyBSA Free [NB21-1022]

Western Blot: Histone H3 [Dimethyl Lys4] AntibodyBSA Free [NB21-1022]

Western Blot: Histone H3 [Dimethyl Lys4] Antibody [NB21-1022] - Western blot analysis of H3 K4me2 in HeLa histone preps at 1:1000. Observed molecular weight is ~15 kDa.
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Applications for Histone H3 [Dimethyl Lys4] Antibody - BSA Free

Application
Recommended Usage

Chromatin Immunoprecipitation (ChIP)

2-5 ug per million cells

Dot Blot

1:1000

Immunocytochemistry/ Immunofluorescence

1:50

Immunohistochemistry

reported in scientific literature (PMID 33872374)

Western Blot

1:500
Application Notes
In Western Blot, a band is seen ~15kDa in HeLa histone preps and C. elegans embryo lysate.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 1 review rated 5 using NB21-1022 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS and 30% Glycerol

Format

BSA Free

Preservative

0.05% Sodium Azide

Concentration

0.70 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, H3K4Me2

Entrez Gene IDs

126961 (Human); 260423 (Mouse)

Gene Symbol

H3C14

UniProt

Q71DI3 (Human), P84228 (Mouse)

Additional Histone H3 Products

Product Documents for Histone H3 [Dimethyl Lys4] Antibody - BSA Free

Product Datasheets

Certificate of Analysis

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Product Specific Notices for Histone H3 [Dimethyl Lys4] Antibody - BSA Free

Epi-Plus antibody production in collaboration with Rockland Immunochemicals Inc.

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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