ICC/IF image of ab49652 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab49652, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Western blot - Anti-Calpain 1 antibody [15C10] (ab49652)Image courtesy of an anonymous Abreview.
All lanes : Anti-Calpain 1 antibody [15C10] (ab49652) at 1/1000 dilution
Lane 1 : Nuclear tissue lysate prepared from rat heart, left ventricle Lane 2 : Nuclear tissue lysate prepared from rat heart, left ventricle
Lysates/proteins at 30 µg per lane.
Secondary HRP-conjugated donkey anti-mouse polyclonal at 1/10000 dilution Developed using the ECL technique
Predicted band size : 80 kDa Observed band size : 80 kDa
Overlay histogram showing HeLa cells stained with ab49652 (red line). The cells were fixed with 80% methanol (5 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 (ab49652, 1µg/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 IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line). Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
References for Anti-Calpain 1 antibody [15C10] (ab49652)
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