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

Product name

Anti-Aurora B antibody [EPR25417-73]
See all Aurora B primary antibodies
Description

Rabbit monoclonal [EPR25417-73] to Aurora B
Host species

Rabbit
Tested applications

Suitable for: WB, IHC-P, ICC, IP, Flow Cyt (Intra)more details
Species reactivity

Reacts with: Mouse, Rat
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: L-929 treated with 100ng/ml nocodazole for 20 hours whole, C6, His-tagged mouse Aurora B recombinant protein (aa13-261) protein lysates. IHC-P: Mouse testis, Mouse embryo 12.5 dpc and Mouse colon tissues. ICC: NIH/3T3, L-929 cells. Flow Cyt (intra): NIH/3T3 cell. IP: NIH/3T3 cells.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR25417-73
Isotype

IgG
Research areas

Alternative Versions
- Anti-Aurora B antibody [EPR25417-73] - BSA and Azide free (ab287965)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Related Products
- VeriBlot for IP Detection Reagent (HRP) (ab131366)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab287960 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
WB		1/1000. Predicted molecular weight: 39 kDa.
IHC-P		1/300. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC		1/50.
IP		1/30.
Flow Cyt (Intra)		1/500.

Notes
WB 1/1000. Predicted molecular weight: 39 kDa.
IHC-P 1/300. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC 1/50.
IP 1/30.
Flow Cyt (Intra) 1/500.

Function

May be directly involved in regulating the cleavage of polar spindle microtubules and is a key regulator for the onset of cytokinesis during mitosis. Component of the chromosomal passenger complex (CPC), a complex that acts as a key regulator of mitosis. The CPC complex has essential functions at the centromere in ensuring correct chromosome alignment and segregation and is required for chromatin-induced microtubule stabilization and spindle assembly. Phosphorylates 'Ser-10' and 'Ser-28' of histone H3 during mitosis. Required for kinetochore localization of BUB1 and SGOL1. Interacts with INCENP.
Tissue specificity

High level expression seen in the thymus. It is also expressed in the spleen, lung, testis, colon, placenta and fetal liver. Expressed during S and G2/M phase and expression is up-regulated in cancer cells during M phase.
Involvement in disease

Note=Disruptive regulation of expression is a possibile mechanism of the perturbation of chromosomal integrity in cancer cells through its dominant-negative effect on cytokinesis.
Sequence similarities

Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. Aurora subfamily.
Contains 1 protein kinase domain.
Post-translational
modifications

Ubiquitinated by different BCR (BTB-CUL3-RBX1) E3 ubiquitin ligase complexes. Ubiquitinated by the BCR(KLHL9-KLHL13) E3 ubiquitin ligase complex, ubiquitination leads to removal from mitotic chromosomes and is required for cytokinesis. During anaphase, the BCR(KLHL21) E3 ubiquitin ligase complex recruits the CPC complex from chromosomes to the spindle midzone and mediates the ubiquitination of AURKB. Ubiquitination of AURKB by BCR(KLHL21) E3 ubiquitin ligase complex may not lead to its degradation by the proteasome.
Cellular localization

Nucleus. Chromosome. Chromosome > centromere. Cytoplasm > cytoskeleton > spindle. Localizes on chromosome arms and inner centromeres from prophase through metaphase and then transferring to the spindle midzone and midbody from anaphase through cytokinesis. Colocalized with gamma tubulin in the mid-body.
Target information above from: UniProt accession Q96GD4 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 20877 Mouse
- Entrez Gene: 114592 Rat
- SwissProt: O70126 Mouse
- SwissProt: O55099 Rat
- Unigene: 3488 Mouse
- Unigene: 10865 Rat
Alternative names
- AIK2 antibody
- AIM-1 antibody
- AIM1 antibody
- ARK-2 antibody
- ARK2 antibody
- AurB antibody
- AURKB antibody
- AURKB_HUMAN antibody
- Aurora 1 antibody
- Aurora and Ipl1 like midbody associated protein 1 antibody
- Aurora kinase B antibody
- Aurora related kinase 2 antibody
- Aurora- and Ipl1-like midbody-associated protein 1 antibody
- Aurora-B antibody
- Aurora-related kinase 2 antibody
- Aurora/IPL1 related kinase 2 antibody
- Aurora/IPL1-related kinase 2 antibody
- IPL1 antibody
- PPP1R48 antibody
- Protein phosphatase 1 regulatory subunit 48 antibody
- Serine/theronine kinase 12 antibody
- Serine/threonine protein kinase 12 antibody
- Serine/threonine-protein kinase 12 antibody
- Serine/threonine-protein kinase aurora-B antibody
- STK-1 antibody
- STK1 antibody
- STK12 antibody
- STK5 antibody
see all

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

All lanes : Anti-Aurora B antibody [EPR25417-73] (ab287960) at 1/1000 dilution

Lane 1 : L-929 ( mouse connective tissue fibroblast) whole cell lysate
Lane 2 : L-929 treated with 100ng/ml nocodazole for 20 hours whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution

Predicted band size: 39 kDa

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

Exposure time: 3 minutes
Immunocytochemistry - 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.
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
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.
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.
Western blot - Anti-Aurora B antibody [EPR25417-73] (ab287960)

All lanes : Anti-Aurora B antibody [EPR25417-73] (ab287960) at 1/1000 dilution

Lane 1 : His-tagged mouse Aurora B recombinant protein (aa13-261) 10 ng
Lane 2 : His-tagged mouse Aurora C recombinant protein (aa5-237) 10 ng
Lane 3 : GST-tagged mouse Aurora A recombinant protein (fl-length) 10ng

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

Predicted band size: 39 kDa

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

Exposure time: 5.5 seconds
Immunocytochemistry - 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.
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
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
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
Western blot - Anti-Aurora B antibody [EPR25417-73] (ab287960)

Anti-Aurora B antibody [EPR25417-73] (ab287960) at 1/1000 dilution + C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution

Predicted band size: 39 kDa

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

This blot was developed using a higher sensitivity ECL substrate.

Exposure time: 3 minutes
Anti-Aurora B antibody [EPR25417-73] (ab287960)

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

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Key features and details

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Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Involvement in disease

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (0)

Customer reviews and Q&As

Post-translational
modifications