Background-subtracted data values (mean +/- SD) are graphed.

Background-subtracted data values (mean +/- SD) are graphed. 

Background-subtracted data values (mean +/- SD) are graphed. 

Interpolated values of AMPK-alpha 1 are plotted for the indicated cell lines at 100 µg/mL lysate loading. Corresponding Western Blot bands at 40 ug per lane loading are shown for the capture (A) and detector (B) antibodies. Hek293T cells show significantly higher levels of AMPK-alpha 1 both by western blot and by SimpleStep ELISA

Endogenous levels of AMPK-alpha 1 were measured from various cell extracts on western blot using the capture and detector antibodies provided in the kit. Cells were extracted using the protocol outlined in Section 11 and supplemented with SDS sample buffer prior to loading. Samples were loaded as follows: (1) 40 µg Hek293T, (2) 40 µg HeLa, (3) 40 µg MCF7, and (4) 40 µg HepG2. Samples 2-5 were blotted with the detector antibody, and samples 6-9 were blotted with the capture antibody. Blots were performed under reducing conditions. Membranes were blocked with 5% Milk in TBS + 0.1% Tween-20 (TBST) for 1 hour at room temperature. Primary antibodies were incubated in 1% Milk in TBST overnight. Secondary antibodies (goat anti-rabbit HRP for detector and goat anti-mouse HRP for capture) were incubated in 1X Blocking Buffer (ab126587) for 2 hours at room temperature.