Recombinant Anti-Aurora A antibody [EPR5026] - Centrosome Marker (ab108353)

Lane 1: Neuro2a (Mouse neuroblastoma neuroblast) whole cell lysate, 10µg
Lane 2: Neuro2a cell lysate 350µg and ab109518 2µg
Lane 3: Neuro2a cell lysate, 350µg and rabbit IgG (ab172730) , 2µg

Purified ab108353 immunoprecipitating Aurora A in Neuro2a cell lysates. Primary antibody was used at a 1/60 dilution (20 µg/ml).  For western blotting, ab108353 at 1/500 and VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Blocking and diluting buffer used: 5% NFDM/TBST.

Lane 1: HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate 10µg
Lane 2: HepG2 cell lysate 350µg and ab109518 2µg
Lane 3:  HepG2 cell lysate 350µg and rabbit IgG (ab172730) , 2µg

Purified ab108353 immunoprecipitating Aurora A in Neuro2a cell lysates. Primary antibody was used at a 1/60 dilution (20 µg/ml).  For western blotting, ab108353 at 1/500 and VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Blocking and diluting buffer used: 5% NFDM/TBST.

Overlay histogram showing HeLa cells stained with ab108353 (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 (ab108353, 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.