All batches of ab174566 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 - asymmetric di methyl R17 peptide (ab16935) and Histone H3 - symmetric di methyl R17 peptide (ab32948).ab14663 - Histone H3 - unmodifiedab16935 - Histone H3 - asymmetric di methyl R71ab32948 - Histone H3 - symmetric di methyl R17
ICC/IF image of ab174566 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 ab174566 at 0.2µg/ml overnight at +4°C. The secondary antibody (green) was a goat anti-rabbit DyLight® 488 (ab96899) IgG (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.
Anti-Histone H3 (asymmetric di methyl R17, symmetric di methyl R17) antibody (ab174566) at 1 µg/ml + HeLa Histone Preparation Nuclear Lysate (ab28930) at 2.5 µgSecondaryGoat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) (ab65484) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.