Competitive inhibition ELISA using ab134901. 50µl of cAMP-SPDP-BSA was coated in 96-wells. After 1% BSA blocking, 25 µl cAMP, AMP, ADP, ATP, cGMP, GMP, GDP, GTP, cIMP, cCMP, Adenosine, H2O and 25 µl of ab134901 were added. HRP conjugated goat anti-rabbit IgG-Fc-HR antibody was used to develop the color.
Competitive ELISA assay using ab134901. 50 µl cAMP-SPDP-BSA was coated in 96-wells. After 1% BSA blocking, serial dilution of ab134901, 25 µl of cAMP, and H2O (negative control) were added. HRP conjugated goat anti-rabbit IgG-Fc antibody was used to develop the color.
ab134901 staining cAMP in SK-N-SH cells treated with (R)-(+)-Methanandamide (ethanol solution) (ab120361), by ICC/IF. Increase in cAMP expression correlates with increased concentration of (R)-(+)-Methanandamide (ethanol solution), as described in literature.The cells were incubated at 37°C for 10 minutes in media containing different concentrations of ab120361 ((R)-(+)-Methanandamide (ethanol solution)) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab134901 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody.