ANTI-TFAM. CLONE 20G2C12 25 Y56 (C005B-451129)

General description

Histone H3.3 (UniProt: P84243; also known as Histone H3, H3.3) is encoded by the H3-3A (also known as H3.3A, H3F3, H3F3A) gene (Gene ID: 3020; 3021) in human. Histones are basic nuclear proteins that are responsible for the nucleosome structure of chromatin in eukaryotes. Two molecules of each of the four core histones (H2A, H2B, H3, and H4) form an octamer, around which DNA is wrapped in repeating units, called nucleosomes, which limits DNA accessibility to the cellular machineries, which require DNA as a template. Histone H3 has two main variants, H3.1 and H3.3, which show different genomic localization patterns in animals. Histone H3.1 serves as the canonical histone, which is incorporated during DNA replication. Histone H3.3 is a highly conserved variant form of Histone H3, which replaces conventional H3 in a wide range of nucleosomes in active genes. It constitutes the predominant form of histone H3 in non-dividing cells and is incorporated into chromatin independently of DNA synthesis. It is associated with actively expressed genes in both animals and plants. It is predominantly enriched near transcription end sites (TES) of genes and positively associated with transcription. Protein arginine methyltransferase 6 (PRMT6) is known to catalyze asymmetric dimethylation of histone H3 at arginine 2 (H3R2me2a) and this asymmetric demethylation is reported to associate with silent genes. H3R2me2a contributes to gene repression by blocking the catalytic activity of the H3K4 methyltransferase KMT2A and counteracting the formation of H3K4 trimethylation. Transcriptional repression by PRMT6 and H3R2me2a is reported to be relevant for pluripotency and differentiation processes. It is also required for the maintenance of fully differentiated cells in various tissues. PRMT6-dependent H3R2me2a is also detected at active genes, both at promoter and enhancer sites. It contributes to gene repression at promoter and gene activation at enhancer sites. Loss of H3R2me2a from promoter sites is reported to result in enhanced KMT2A binding and H3K4me3 deposition along with increased target gene transcription. At enhancers, H3R2me2a peaks colocalize with the active enhancer marks H3K4me1 and H3K27ac. (Ref.: Bouchard, C., et al. (2018). Cell Reports 24(12); 3339-3352; Chow, C-M., et al. (2005). EMBO Rep. 6(4); 354-360; Tagami, H., et al. (2004). Cell. 116(1); 51-61).

Specificity

Clone 18F3 is a rat monoclonal antibody that detects Histone H3.3 dimethylated on arginine 2.

Immunogen

Ovalbumin-conjugated linear peptide corresponding to 8 amino acids from the N-terminal region of human Histone H3.3 dimethylated on arginine 2.

Application

Quality Control Testing

Evaluated by Dot Blot analysis with Histone H3 peptide dimethylated on arginine 2 (H3R2me2a).

Tested Applications

DOT Blot Analysis: 1:1,000 dilution of this antibody detected 0.05 to 1.0 g of Histone H3 peptide dimethylated on arginine 2 (H3R2me2a).

Chromatin Immunoprecipitation Analysis (ChIP): A representative lot detected Histone H3R2me2a in Chromatin Immunoprecipitation application (Bouchard, C., et al. (2018). Cell Rep. 24(12):3339-3352).

Dot Blot Analysis: A representative lot detected Histone H3R2me2a in Dot Blot application (Bouchard, C., et al. (2018). Cell Rep. 24(12):3339-3352).

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.

Physical form

Purified rat monoclonal antibody IgG2c in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Reconstitution

1.0 mg/mL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Storage and Stability

Recommended storage: +2°C to +8°C.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.