Anti-muscle Actin antibody [EP184E] (ab46805) at 1/1000 dilution + A431 cell lysate at 10ug/lane

Secondary
Goat anti-rabbit, HRP labelled. at 1/2000 dilution

Predicted band size: 42 kDa
Observed band size: 42 kDa

Immunofluorescent staining of HeLa cells using ab46805 at a 1/250 dilution.

Immunohistochemical staining of paraffin-embedded human cardiac muscle using anti-Actin ab46805 at a 1/100 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Overlay histogram showing HeLa cells stained with ab46805 (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 (ab46805, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monclonal) (1µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

Fluorescent immunohistochemical analysis of paraffin-embedded human smooth muscle tissue using ab46805. Green-specific fluorescent staining.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.