APC Anti-MelanA antibody [EPR20380]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - APC Anti-MelanA antibody [EPR20380] (AB225498)

Flow cytometry overlay histogram showing MeWo positive cells (right) and negative HEK293 cells (right) stained with ab225498 (red line). The cells were fixed with 4 % formaldehyde (10 min) and then permeabilized with 90% methanol. The cells were then incubated in 1x PBS containing 10 % normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab225498) (1x 10⁶ in 100 μl at 1 μg/ml (1/500)) for 30 min at 22°C.

Isotype control antibody (black line) was used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5000 events were collected using a 40 mW Red laser (638nm) and 660/10 bandpass filter.

• ICC

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Immunocytochemistry - APC Anti-MelanA antibody [EPR20380] (AB225498)

ab225498 staining MelanA in B16-F10 cells, with negative expression in HEK293 cells. The cells were fixed with 100% methanol (5 min), permeabilised with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab225498 at 0.2 μg/ml (shown in Red). ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488) (shown in green) was used at 2 μg/ml for structural counterstaining.

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.