Western blot against tagged recombinant protein immunogen using ab57484 ADAM17 antibody at 1ug/ml.

ab57484 was used to detect mouse ADAM17 overexpressed in Hela cells. The transfected cells were incubated with ab57484 (10 µg/ml ) for 1 hour at  22°C. For further details please refer to the Abreview.

Overlay histogram showing HeLa cells stained with ab57484 (red line). The cells were fixed with methanol (5 min) and 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 (ab57484, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in HeLa cells fixed with 4% paraformaldehyde (10 min) used under the same conditions.Please note that Abcam do not have data for use of this antibody on non-fixed cells. We welcome any customer feedback.

Anti-ADAM17 antibody (ab57484) at 1/1000 dilution + whole cell lysate prepared from MCF7 cells at 100000 cellsSecondaryHRP conjugated sheep anti-mouse polyclonal at 1/2000 dilutiondeveloped using the ECL techniqueObserved band size : 50 kDa (why is the actual band size different from the predicted?)Exposure time : 1 minuteImage courtesy of an anonymous Abreview.See Abreview