ANTI-RAD51 ANTIBODY (C15-1662-289)

General description

Histone H3.1t (UniProt: Q16695; also known as H3/t, H3t, H3/g) is encoded by the HIST3H3 (also known as H3FT) gene (Gene ID: 8290) in human. Histones are highly conserved basic nuclear proteins that are responsible for the nucleosome structure of chromatin in eukaryotes. They play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling. 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 that require DNA as a template. Histone H3 features a main globular domain and a long N-terminal tail, which protrudes from the globular nucleosome core and can undergo several different types of epigenetic modifications that influence cellular processes. The amino-terminal tails of histone proteins are subject to posttranslational modifications, including acetylation and methylation, which recruit downstream regulatory factors, influence chromatin structure, and are critical determinants of transcription. Acetylation of histone H3 occurs at several different lysine positions in the histone tail and is performed by a family histone acetyltransferases. Acetylation is generally associated with transcriptional activity and methylation of lysine and arginine residues can either activate or repress depending on the residue modified. There is a significant correlation between acetylation of histones H3 in promoter regions and transcriptional activity. The presence of H3K4me3 has been correlated with transcriptionally active promoters of genes and its levels positively correlate with gene expression levels. H3K4me3 has been implicated in a number of nuclear processes, including Pol II-mediated transcription, pre-mRNA splicing, DNA recombination, and DNA repair. Histaminylation of glutamine 5 on histone H3 (H3Q5Histaminyl) is recently identified as a permissive posttranslational modification. (Ref.: Zhao, S., et al. (2021). Proc. Natl. Acad. Sci. USA. 118(6):e2016742118; Diehl , KL., and Muir, TW. (2020). Nat. Chem. Biol. 16; 620-629; Bernstein, BE., et al. (2002). Proc. Natl. Acad. Sci. USA. 99(13);8695-8700).

Specificity

This rabbit polyclonal antibody detects Histone H3.1 histaminylated on glutamine 5.

Immunogen

KLH-conjugated linear peptide corresponding to 10 amino acids from the N-terminal region of human Histone H3.1 with histaminylated glutamine 5 (without the initiator methionine).

Application

Quality Control Testing

Evaluated by DOT Blot analysis with H3Q5Histaminyl peptide

Dot Blot Analysis: A 1:1,000 dilution of this antibody detected H3Q5Histaminyl peptide, but not the unmodified H3Q5 peptide.

Tested Applications

Western Blotting Analysis: A representative lot detected H3Q5histaminyl in lysate from Transglutaminase 2 (TGM2) overexpressing HEK293T cells treated with Histamine, but not in untreated cells or in cells treated with Serotonin or Dopamine. (Data courtesy of Dr. Ian Maze′s Lab, Icahn School of Medicine at Mount Sinai, New York).

Western Blotting Analysis: A 1:1,000 dilution of this antibody detected Histone H3Q5histaminyl in lysates from HEK293T cells over expressing TGM2 and treated with histamine, but not in untreated cells.

Dot Blot Analysis: A 1:1,000 dilution of a representative lot of this antibody detected H3Q5histaminyl peptide, but not serotonin or dopamine modified peptides. (Data courtesy of Dr. Ian Maze s Lab, Icahn School of Medicine at Mount Sinai, New York).

Activity Assay (In vitro): A representative lot of this antibody detected the recombinant Histone H3.3 histaminylated on glutamine 5 following in vitro transamidation reaction in the presence of histamine. (Data courtesy of Dr. Ian Maze Lab, Icahn School of Medicine at Mount Sinai, New York).

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

Anti-H3Q5histaminyl, Cat. No. ABE2578, is a rabbit polyclonal antibody that detects Histone H3.1 histaminylated on glutamine 5 and is tested for use in Activity Assay (in vitro), Dot Blot, and Western Blotting,

Physical form

Purified rabbit polyclonal antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide

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.