Anti-Aurora B antibody [EP1009Y] - BSA and Azide free

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Aurora B antibody [EP1009Y] - BSA and Azide free (AB239837)

This data was developed using ab45145, the same antibody clone in a different buffer formulation. Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Aurora B with Purified ab45145 at 1/300 dilution (0.1 μg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488 ,ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Left). Unlabeled control - /.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Aurora B antibody [EP1009Y] - BSA and Azide free (AB239837)

This data was developed using ab45145, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Aurora B with Purified ab45145 at 1 : 50 dilution (5.4 μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1 : 200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor® 488,ab150077) was used as the secondary antibody at 1 : 1000 (2 μg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aurora B antibody [EP1009Y] - BSA and Azide free (AB239837)

This data was developed using ab45145, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human endometrium carcinoma tissue sections labeling Aurora B with purified ab45145 at 1/200 dilution (1.35 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• IP

Lab

Immunoprecipitation - Anti-Aurora B antibody [EP1009Y] - BSA and Azide free (AB239837)

This data was developed using ab45145, the same antibody clone in a different buffer formulation.
Purified ab45145 at 1/20 dilution (1μg) immunoprecipitating Aurora B in HeLa whole cell lysate.
Lane 1 (input) : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10μg
Lane 2 (+) : ab45145 + HeLa whole cell lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab45145 in HeLa whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM/TBST.
Observed band size : 39 kDa

All lanes:

Immunoprecipitation - Anti-Aurora B antibody [EP1009Y] (<a href='/en-us/products/primary-antibodies/aurora-b-antibody-ep1009y-ab45145'>ab45145</a>)

Predicted band size: 39 kDa

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• OI-RD Scanning

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OI-RD Scanning - Anti-Aurora B antibody [EP1009Y] - BSA and Azide free (AB239837)

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.