Application

Monoclonal Anti-phospho-Histone H3 (pSer28) antibody is suitable for immunoblotting (approx. 15 kDa), immunocytochemistry (3.7% formaldehyde-methanol fixation) and flow cytometry applications.

Biochem/physiol Actions

Mitogenic stimulation, oncogenic transformation, or induction of oncogenic ras expression are accompanied with increase in serine-10 phosphorylation of the H3 N-terminal domain. It has been shown that phosphorylated H3 is associated with c-fos and c-myc genes in stimulated cells. Phosphorylation of H3, at both serine-10 and -28, coincides with the induction of mitotic chromosome condensation.H3 phosphorylation may contribute to proto-oncogene induction by modulating chromatin structure and releasing blocks in elongation. H3 dephosphorylation occurs quite rapidly after mitosis and serine-10/28 reo main unphosphorylated throughout the remainder of interphase. PP1 has been identified as the H3 phosphatase.

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 H1 and the N-terminal tail of histone-3 (H3) have key roles in the folding and inter-association of the chromatin fibers. Two currently known phosphorylation sites are present in the N-terminus of H3; serine-10 and serine-28. Mammals contain three main classes of histone H3 variants: the replicative histones (H3.1 and H3.2), the replacement histone (H3.3) and the centromeric histone (Cenp-A). Histone H3 is located on human chromosome 6p22.

Immunogen

Synthetic peptide corresponding to amino acids of human histone H3.

Physical form

Solution in 0.01 M phosphate buffered saline, pH 7.4, containing 1% BSA and 15 mM sodium azide.

Preparation Note

Purified from culture supernatant of hybridoma cells grown in a bioreactor

Specificity

The antibody does not detect the unphosphorylated epitope. It detects the histone molecule at the onset of mitosis (prophase, metaphase, weaker at the beginning of anaphase, but not during late anaphase).

Monoclonal Anti-Phospho-Histone H3 (pSer28) reacts specifically with histone H3 phosphorylated at Ser28 near its N-terminus, and does not detect the unphosphorylated epitope. It detects the phosphorylated histone molecule at the onset of mitosis (prophase, metaphase and weaker at the beginning of anaphase), but not during late anaphase. Reactivity has been observed with human, bovine, hamster and mouse.

Storage and Stability

For continuous use, store at 2 to 8 °C for up to one month. For extended storage, freeze in working aliquots. Repeated freezing and thawing is not recommended. Storage in "frost-free" freezers is not recommended. If slight turbidity occurs upon prolonged storage, clarify the solution by centrifugation before use. Working dilution samples should be discarded if not used within 12 hours.