ab33315 staining human fibrosarcoma cells by ICC/IF. Cells were PFA fixed and blocked with 100% Image-iT^TM signal enhancer for 30 minutes at 20°C. The primary antibody was used at 10µg/ml concentration and incubated with sample for 1 hour at 25°C. Alexa Fluor®555 conjugated donkey monoclonal to mouse (H+L) was used as the secondary antibody, diluted 1/500.  

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Anti-CD151 antibody [11G5a] (ab33315) at 1/1000 dilution + HT1080 (human fibrosarcoma cell line) whole cell lysate

Secondary
Sheep anti-mouse IgG HRP at 1/2000 dilution

Developed using the ECL technique.

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ICC/IF image of ab33315 stained HT 1080 cells. The cells were 4% formaldehyde fixed (10 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 (ab33315, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96789, 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.