ICC/IF image of ab130940 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab130940, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, a goat anti-rabbit DyLight® 488 (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.This antibody also gave a positive result in 100% methanol fixed (5 min) Hek293, HepG2 and MCF7 cells at 1µg/ml.
Anti-Histone H3 (phospho T3) antibody (ab130940) at 1 µg/ml + Calf Thymus Histone Preparation Nuclear Lysate at 0.25 µgSecondaryGoat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
All batches of ab130940 are tested in Peptide Array against peptides to different Histone H3 modifications. Six dilutions of each peptide are printed on to the Peptide Array in triplicate and results are averaged before being plotted on to a graph. Results show strong binding to Histone H3 - phospho T3 peptide (ab154294), indicating that this antibody specifically recognises the Histone H3 - phospho T3 modification.ab154294 - Histone H3 - phospho T3ab7228 - Histone H3 - unmodifiedab14103 - Histone H3 - phospho T6ab24444 - Histone H3 - phospho T11