Anti-Aurora A antibody [EPR5026] - Centrosome Marker

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Aurora A antibody [EPR5026] - Centrosome Marker (AB108353)

Immunofluorescent staining of HeLa cells using ab108353 at 1/100

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Aurora A antibody [EPR5026] - Centrosome Marker (AB108353)

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 >5000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

• IP

Lab

Immunoprecipitation - Anti-Aurora A antibody [EPR5026] - Centrosome Marker (AB108353)

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.

All lanes:

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

Predicted band size: 45 kDa

false

• IP

Lab

Immunoprecipitation - 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.

All lanes:

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

Predicted band size: 45 kDa

false

• WB

Unknown

Western blot - Anti-Aurora A antibody [EPR5026] - Centrosome Marker (AB108353)

All lanes:

Western blot - Anti-Aurora A antibody [EPR5026] - Centrosome Marker (ab108353) at 1/1000 dilution

Lane 1:

BXPC-3 cell lysate at 10 µg

Lane 2:

LnCaP cell lysate at 10 µg

Lane 3:

SKBR-3 cell lysate at 10 µg

Lane 4:

HepG2 cell lysate at 10 µg

Predicted band size: 45 kDa

false

• WB

Unknown

Western blot - Anti-Aurora A antibody [EPR5026] - Centrosome Marker (AB108353)

All lanes:

Western blot - Anti-Aurora A antibody [EPR5026] - Centrosome Marker (ab108353) at 1/1000 dilution

All lanes:

Neuro-2a cell lysate at 10 µg

Predicted band size: 134 kDa,45 kDa,53 kDa,86 kDa

false

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-Aurora A antibody [EPR5026] - Centrosome Marker (AB108353)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.