ICC/IF image of ab58284 stained MALME-3M 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 ab58284 at 10µg/ml overnight at +4°C. The secondary antibody (pseudo-colored green) was Dylight® 488 goat anti- mouse (ab96879) IgG (H+L) preadsorbed, used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1h at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.
Immunofluorescence of Jurkat cells using ab58284 to stain Tyrosinase antibody diluted to 10 μg/ml.
Overlay histogram showing Malme-3M cells stained with ab126577 (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 (ab126577, 0.1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H+L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
Tyrosinase antibody (ab58284) used in immunohistochemistry at 3ug/ml on formalin fixed and paraffin embedded human liver tissue.
Tyrosinase antibody (ab58284) at 1ug/lane + A-431 cell lysate at 25ug/lane.