Overlay histogram showing HepG2 cells stained with ab118329 (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 followed by the antibody (ab118329, 0.1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HepG2 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
Anti-Retinoid X Receptor alpha antibody (ab118329) at 5 µg/ml + Recombinant immunogen at 0.2 µgdeveloped using the ECL technique
Anti-Retinoid X Receptor alpha antibody (ab118329) at 5 µg/ml + Human kidney tissue lysate at 50 µgdeveloped using the ECL technique
ab118329 at 10 µg/ml as a capture antibody. The detection limit for recombinant tagged Retinoid X Receptor alpha is 0.1 ng/ml.