Histone H3 Antibody (1B1-B2) - Azide and BSA Free

Novus Biologicals | Catalog # NBP2-80770

Novus Biologicals

Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Human, Mouse, Rat

Applications

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Flow Cytometry, Flow (Intracellular), Immunocytochemistry/ Immunofluorescence, Chromatin Immunoprecipitation (ChIP), Single Cell Western

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG3 Kappa Clone # 1B1-B2

Format

Azide and BSA Free
View all Histone H3 Primary Antibodies »

Product Specifications

Immunogen

This Histone H3 antibody (1B1-B2) was raised against synthetic peptide corresponding to the C-terminus of human Histone H3.

Localization

Nucleus. Chromosome.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG3 Kappa

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 Antibody (1B1-B2) - Azide and BSA Free

Western Blot: Histone H3 Antibody (1B1-B2)Azide and BSA Free [NBP2-80770]

Western Blot: Histone H3 Antibody (1B1-B2)Azide and BSA Free [NBP2-80770]

Western Blot: Histone H3 Antibody (1B1-B2) - Azide and BSA Free [NBP2-80770] - Western blot image of Anti-Histone H3 antibody (clone 1B1-B2). Whole cell protein from HeLa (lane 1), NIH-3T3 (lane 2) and PC12 (lane 3) were separated by 4-15% SDS-PAGE and transfered to PVDF membrane. The membrane was probed with purified Anti-Histone H3 antibody at 2 ug/ml and detected with an HRP conjugated anti-mouse secondary using chemiluminescence. Observed molecular weight is ~15 kDa. Image from the standard format of this antibody.
Immunocytochemistry/ Immunofluorescence: Histone H3 Antibody (1B1-B2) - Azide and BSA Free [NBP2-80770]

Immunocytochemistry/ Immunofluorescence: Histone H3 Antibody (1B1-B2) - Azide and BSA Free [NBP2-80770]

Immunocytochemistry/Immunofluorescence: Histone H3 Antibody (1B1-B2) - Azide and BSA Free [NBP2-80770] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton X-100. The cells were incubated with anti-Histone H3 (1B1-B2) NBP2-36468 at a 1:200 dilution overnight at 4C and detected with an anti-mouse DyLight 488 (Green) at a 1:500 dilution. Actin was detected with Phalloidin 568 (Red) at a 1:200 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective. Image from the standard format of this antibody.
Immunohistochemistry: Histone H3 Antibody (1B1-B2) - Azide and BSA Free [NBP2-80770]

Immunohistochemistry: Histone H3 Antibody (1B1-B2) - Azide and BSA Free [NBP2-80770]

Immunohistochemistry: Histone H3 Antibody (1B1-B2) - Azide and BSA Free [NBP2-80770] - Analysis of Histone H3 in human breast cancer using DAB with hematoxylin counterstain. Image from the standard format of this antibody.
Flow Cytometry: Histone H3 Antibody (1B1-B2) - Azide and BSA Free [NBP2-80770]

Flow Cytometry: Histone H3 Antibody (1B1-B2) - Azide and BSA Free [NBP2-80770]

Flow Cytometry: Histone H3 Antibody (1B1-B2) - Azide and BSA Free [NBP2-80770] - Analysis using Alexa Fluor (R) 647 conjugate of NBP2-36468. Jurkat cells stained with Alexa Fluor 647 conjugated Histone H3 antibody (red) or isotype control (blue). Flow cytometry image submitted by a verified customer review.
Chromatin Immunoprecipitation: Histone H3 Antibody (1B1-B2) - Azide and BSA Free [NBP2-80770] - Chromatin from one million formaldehyde cross-linked HeLa cells was precipitated using 2 ug of NBP2-36468 and 25 ul of magnetic IgG beads, using standard ChIP methods. A similar sample containing no antibody was included as a negative control. Immunoprecipitated DNA was quantified using quantitative real-time PCR and SYBR green dye, then normalized to the non-precipitated input chromatin. Representative target genes from active, inactive and heterochromatic regions of the genome show amplification, indicative of the presence of Histone H3. Image from the standard format of this antibody.
Western Blot: Histone H3 Antibody (1B1-B2)Azide and BSA Free [NBP2-80770]

Western Blot: Histone H3 Antibody (1B1-B2)Azide and BSA Free [NBP2-80770]

Western Blot: Histone H3 Antibody (1B1-B2) - Azide and BSA Free [NBP2-80770] - Analysis in Hela cells using siRNA to knockdown Pol II, controls are GAPDH and Histone H3. Theoretical molecular weight of Histone H3 is 15 kDa. WB blot image submitted by a verified customer review. Image from the standard format of this antibody.

Applications for Histone H3 Antibody (1B1-B2) - Azide and BSA Free

Application
Recommended Usage

Immunohistochemistry

1:200-1:500

Immunohistochemistry-Paraffin

1:200-1:500

Single Cell Western

100 ug/ml

Western Blot

1:1000. Use reported by customer review

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Format

Azide and BSA Free

Preservative

No Preservative

Concentration

0.5 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 Antibody (1B1-B2) - Azide and BSA Free

Certificate of Analysis

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Product Specific Notices for Histone H3 Antibody (1B1-B2) - Azide and BSA Free

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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Protocols

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