ELISA using ab16974 at antibody concentrations between 1/100 and 1/10000.The orange line indicates binding to the Histone H4 peptide - mono methyl K20 (ab17043). Binding to the following peptides was either not seen or was very weak:Histone H4 peptide - tri methyl K20 (ab17567) - green line,Histone H4 peptide - di methyl K20 (ab14964) - blue line,Histone H4 peptide - unmodified (ab2622) - red line.This indicates the specificity of ab16794 for mono methyl K20 of Histone H4.
![Overlay histogram showing HeLa cells stained with ab16974 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab16974, 2µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_3/2327_Histone-H4-Primary-antibodies-ab16974-1.jpg)
Overlay histogram showing HeLa cells stained with ab16974 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab16974, 2µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.