All batches of ab129231 are tested in Peptide Array against peptides to different Histone H4 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 H4 - asymmetric di methyl R3 peptide (ab154780), indicating that this antibody specifically recognises the Histone H4 - asymmetric di methyl R3 modification.ab154780 - Histone H4 - asymmetric di methyl R3ab14791 - Histone H4 - symmetric di methyl R3ab17770 - Histone H4 - mono methyl R3ab154467 - Histone H4 - unmodified
Anti-Histone H4 (asymmetric di methyl R3) antibody (ab129231) 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.
ICC/IF image of ab129231 stained MCF-7 cells. The cells were 4% formaldehyde fixed (10 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 ab129231 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 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.