Anti-Hexokinase II antibody (ab76959) at 1/1000 dilution + A-431 cell lysate at 25 µg
Anti-Hexokinase II antibody (ab76959) at 1/2500 dilution + Immunizing peptide at 0.1 µg
Detection limit for recombinant GST tagged HK2 is approximately 0.3ng/ml as a capture antibody.
Overlay histogram showing K562 cells stained with ab76959 (red line). The cells were fixed with 4% paraformaldehyde (10 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. The cells were then incubated with the antibody (ab76959, 0.5µg/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 IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in K562 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.