Anti-Aurora B antibody [EPR25417-73]

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Aurora B antibody [EPR25417-73] (AB287960)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse stomach tissue staining CaMKII beta with ab314897 at 1/5000 dilution, ab315150 anti-Serotonin used at 1/5000 dilution and ab287960 anti-Aurora B used at a 1/300 dilution. Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-CaMKII beta (magenta; Opal^™690), anti-Serotonin (green; Opal^™520) and anti-Aurora B (gray; Opal^™570) on mouse stomach.
Panel B : anti-CaMKII beta staining endocrine cells in mouse stomach.
Panel C : anti-Serotonin staining enterochromaffin cells in mouse stomach.
Panel D : anti-Aurora B staining proliferating cells in mouse stomach.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab314897, ab315150 and ab287960 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal^™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aurora B antibody [EPR25417-73] (AB287960)

Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labelling Aurora B with ab287960 at 1/300 (1.803 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) . Negative control : no staining on mouse lung. The section was incubated with ab287960 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) .

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Aurora B antibody [EPR25417-73] (AB287960)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized L-929 cells labelling Aurora B with ab287960 at 1/50 (10.82 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing positive staining in L-929 cells in M phase is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Aurora B antibody [EPR25417-73] (AB287960)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labelling Aurora B with ab287960 at 1/500 dilution (0.1ug)/ Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control. A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Cells were co-stained with DRAQ5 to differentiate cell cycle phase.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aurora B antibody [EPR25417-73] (AB287960)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labelling Aurora B with ab287960 at 1/300 (1.803 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) . Nuclear staining on mouse colon. The section was incubated with ab287960 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) .

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Aurora B antibody [EPR25417-73] (AB287960)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 cells labelling Aurora B with ab287960 at 1/50 (10.82 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing positive staining in NIH/3T3 cells in M phase is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aurora B antibody [EPR25417-73] (AB287960)

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labelling Aurora B with ab287960 at 1/300 (1.803 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™Polymer Refine Detection) . Nuclear staining on mouse testis. The section was incubated with ab287960 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™Polymer Refine Detection) .

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aurora B antibody [EPR25417-73] (AB287960)

Immunohistochemical analysis of paraffin-embedded Mouse embryo 12.5 dpc tissue labelling Aurora B with ab287960 at 1/300 (1.803 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) . Nuclear staining on mouse embryo 12.5 dpc . The section was incubated with ab287960 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) .

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IP

Supplier Data

Immunoprecipitation - Anti-Aurora B antibody [EPR25417-73] (AB287960)

Aurora B was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 ug with anti aurora b antibody epr2541773 immunoprecipitation nih3t3.jpg at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using anti aurora b antibody epr2541773 immunoprecipitation NIH/3T3 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 ug

Lane 2 : ab287960 IP in NIH/3T3 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab287960 in NIH/3T3 whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 3 minutes

This blot was developed using a higher sensitivity ECL substrate.

All lanes:

Immunoprecipitation - Anti-Aurora B antibody [EPR25417-73] (ab287960)

Predicted band size: 39 kDa

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• WB

Lab

Western blot - Anti-Aurora B antibody [EPR25417-73] (AB287960)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

This blot was developed using a higher sensitivity ECL substrate.

Exposure time : 3 minutes

All lanes:

Western blot - Anti-Aurora B antibody [EPR25417-73] (ab287960) at 1/1000 dilution

All lanes:

C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 39 kDa

false

• WB

Lab

Western blot - Anti-Aurora B antibody [EPR25417-73] (AB287960)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Exposure time : 5.5 seconds

All lanes:

Western blot - Anti-Aurora B antibody [EPR25417-73] (ab287960) at 1/1000 dilution

Lane 1:

His-tagged mouse Aurora B recombinant protein (aa13-261) 10ng

Lane 2:

His-tagged mouse Aurora C recombinant protein (aa5-237) 10ng

Lane 3:

GST-tagged mouse Aurora A recombinant protein (fl-length) 10ng

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 39 kDa

false

• WB

Lab

Western blot - Anti-Aurora B antibody [EPR25417-73] (AB287960)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Exposure time : 3 minutes

All lanes:

Western blot - Anti-Aurora B antibody [EPR25417-73] (ab287960) at 1/1000 dilution

Lane 1:

L-929 ( mouse connective tissue fibroblast) whole cell lysate at 20 µg

Lane 2:

L-929 treated with 100ng/ml nocodazole for 20 hours whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 39 kDa

false