Histone H3 [Trimethyl Lys27] Antibody (BT164) - Chimeric - Azide and BSA Free

Novus Biologicals | Catalog # NBP3-12020

Recombinant Monoclonal Antibody
Novus Biologicals
100% Guarantee

Key Product Details

Species Reactivity

Human

Applications

Immunohistochemistry, Western Blot, ELISA, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Kappa Clone # BT164

Format

Azide and BSA Free
View all Histone H3 Primary Antibodies »

Product Specifications

Immunogen

BT164 is a mice-derrived lupus antibody that was established by selection on apoptotic chromatin.

Modification

Trimethyl Lys 27

Specificity

The epitope of BT164 was mapped in the N-terminal tail of histone H3 (27-KSAPAT-32) and included the apoptosis-induced trimethylation of K27.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG Kappa

Scientific Data Images

Western Blot: Histone H3 [Trimethyl Lys27] Antibody (BT164) - Chimeric [NBP3-12020] - Nuclear lysate of HeLa(A) (0.0003ug/ml), Jurkat(B) (0.0003ug/ml) and K562(C) (0.01ug/ml) (35ug protein in RIPA buffer) were resolved on a SDS PAGE gel and blots were probed with the chimeric rabbit version of BT164 (NBP3-12020) before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.
Immunocytochemistry/Immunofluorescence: Histone H3 [Trimethyl Lys27] Antibody (BT164) - Chimeric [NBP3-12020] - Immunofluorescence analysis of paraformaldehyde fixed HeLa cells on Shi-fix(TM) coverslips stained with the chimeric rabbit IgG version of BT164 (NBP3-12020) at 10 ug/ml for 1h followed by Alexa Fluor(R) 488 secondary antibody (2 ug/ml), showing membrane staining. The nuclear stain is DAPI (blue) and the actin stain is phalloidin (red). Panels show from left-right, top-bottom NBP3-12020, DAPI, merged channels and an isotype control. The isotype control was an unknown specificity antibody followed by staining with Alexa Fluor(R) 488 secondary antibody.
Histone H3 [Trimethyl Lys27] Antibody (BT164) - Chimeric - Azide and BSA Free

Flow Cytometry: Histone H3 [Trimethyl Lys27] Antibody (BT164) - Chimeric - Azide and BSA Free [NBP3-12020] -

Flow Cytometry: Histone H3 [Trimethyl Lys27] Antibody (BT164) - Chimeric - Azide and BSA Free [NBP3-12020] - HeLa cells were fixed using 2% PFA and stained with anti-unknown specificity antibody (isotype control, black line) or NBP3-12020 (blue line) at a dilution of 1:100 for 1h at RT. After washing, the bound antibody was detected using a goat anti-rabbit IgG AlexaFluor 488 antibody at a dilution of 1:1000 and cells analyzed using a FACSCanto flow-cytometer.

Applications

Application
Recommended Usage

ELISA

Optimal dilutions of this antibody should be experimentally determined.

Immunocytochemistry/ Immunofluorescence

Optimal dilutions of this antibody should be experimentally determined.

Immunohistochemistry

Optimal dilutions of this antibody should be experimentally determined.

Immunoprecipitation

Optimal dilutions of this antibody should be experimentally determined.

Western Blot

Optimal dilutions of this antibody should be experimentally determined.
Application Notes
This chimeric rabbit antibody was made using the variable domain sequences of the original Mouse IgG2b format, for improved compatibility with existing reagents, assays and techniques.

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

PBS

Format

Azide and BSA Free

Preservative

0.02% Proclin 300

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 for up to 3 months. For longer storage, aliquot and store at -20C.

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

Certificate of Analysis

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

Product Specific Notices

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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