The rabbit-derived antibody is purified with protein A-conjugated agarose followed by phosphorylated histone H3 (Thr80)peptide affinity chromatography. It specifically recognizes histone H3 with threonine phosphorylation at Thr80.
ELISA, Dot blot, Western blot.
Recommended antibody dilution: WB: 1:2000
Recommended antibody dilution
For Western blotting, incubate ?membrane with diluted antibody in 5% nonfat milk, 1 x TBS, 0.1% Tween-20 for ?two hours at room temperature with gentle shaking.??
Use at an assay dependent concentration. Optimal ?dilutions/concentrations should be determined by the end user.??
A.Dot blotting analysis?on indicated amount of peptides using histone H3T80ph rabbit pAb. The list of peptides is included in the table on the right.
B.Western blotting analysis?on 30 μg of crude proteins from HeLa whole cell lysates with (right) or without (left) treatment of Okadaic acid (100 nM, 60min) and calyculin A (CA,100 nM, 1hr) using phospho-histone H3 (Thr80) rabbit pAb (1: 2000).
C.Western blotting analysis?on 30 μg of crude proteins from mouse brain, mouse testis whole tissue lysates and MCF-7 whole cell lysates using phospho-histone H3 (Thr80) rabbit pAb (1: 2000).
Histones are subjected to a variety of enzyme catalyzed modifications, including acetylation, methylation, ubiquitylation and newly discovered succinylation, crotonylation and 2-hydroxyisobutyrylation, etc. These modifications have a direct effect on the accessibility of chromatin to transcription factors and, therefore, on gene expression. Histone H3 phosphorylation is specifically phosphorylated during both mitosis and meiosis that is believed to be mainly involved in transcriptional activation requiring chromatin fiber decondensation, and chromosome compaction during cell division. Histone H3 phosphorylation may initiate at different phases of the cell division in different organisms, but metaphase chromosomes are always found to be heavily phosphorylated.
Storage & Stability
The antibody is kept in PBS with 50% glycerol and 0.01% sodium azide. Upon receipt, please centrifuge the antibody at 12,000 x g for 20 seconds and store the antibody at -20°C. Avoid repeated freeze/thaw cycles. Stable for 12 months from date of receipt. Leave the antibody at room temperature for 2 minutes and gently mixed using pipette before usage.