Analysis Note

ControlUntreated and Colcemid-treated HeLa cell lysates, and in Histone H3 recombinant protein

Application

Detect Histone H3 using this rabbit polyclonal antibody, Anti-phospho Histone H3.3 Antibody (Thr32) validated for use in western blotting, IP & Dot Blot.

Research Sub CategoryHistones

Research CategoryEpigenetics & Nuclear Function

Dot Blot (Specificity) Analysis: 0.2 µg/mL of this antibody detected phospho Histone H3.3 (Thr32), but not non-phospho Histone H3 (Thr32), or other peptides corresponding to modified histones.

Immunoprecipitation Analysis: A representative lot detected phospho Histone H3.3 (Thr32) in IP.

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.

General description

Histone H3 is one of the five main histone proteins involved in the structure of chromatin in eukaryotic cells. Featuring a main globular domain and a long N-terminal tail, H3 is involved with the structure of the nucleosomes of the ′beads on a string′ structure. The N-terminal tail of histone H3 protrudes from the globular nucleosome core and can undergo several different types of epigenetic modifications that influence cellular processes. These modifications include the covalent attachment of methyl or acetyl groups to lysine and arginine amino acids and the phosphorylation of serine or threonine. Histone H3 variants (H3.1, H3.2 and H3.3) have been implicated in the epigenetic memory of cellular state. Genome-wide patterns of H3.3 are dependent on amino acid sequence and change with cellular differentiation at developmentally regulated loci.

Immunogen

KLH-conjugated linear peptide corresponding to a region near the N-terminus of human Histone H3.3 phosphorylated at Thr32.

Epitope: Near N-terminus

Other Notes

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

Physical form

Affinity purified

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

Quality

Evaluated by Western Blotting in untreated and Colcemid-treated HeLa cell lysates, and in Histone H3 recombinant protein.

Western Blotting Analysis: 0.5 µg/mL of this antibody detected phospho Histone H3.3 (Thr32) in 10 µg of Colcemid-treated HeLa cell lysate.

Specificity

This antibody recognizes a region near the N-terminus of Histone H3.3 phosphorylated at Thr32. This antibody demonstrates specificity for phospho Histone H3.3 (Thr32), but does not cross react with non-phospho Histone H3.3 (Thr32) or any other Histone proteins.

Storage and Stability

Stable for 1 year at 2-8°C from date of receipt.

Target description

~17 kDa observed. Uncharacterized band(s) may be observed in some cell lysates.