Anti-Caspase-6/CASP-6 antibody [EP1325Y] (ab52951) at 1/20000 dilution + MCF-7 cell lysate at 10 µg

Secondary
goat anti-rabbit HRP at 1/2000 dilution

Predicted band size: 34 kDa
Observed band size: 34 kDa

ab52951 at 1:100 dilution staining Caspase-6/CASP-6 in human breast carcinoma by Immunohistochemistry, Paraffin embedded tissue.

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

ab52951 at 1:100 dilution staining Caspase-6/CASP-6 in HeLa cells by Immunofluorescence.

Overlay histogram showing Hela cells stained with ab52951 (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 (ab52951, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/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. 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.