Anti-Aurora B antibody ab2254 is a rabbit polyclonal antibody that is used in Aurora B western blotting, IHC and immunofluorescence. Suitable for human, mouse and rat samples.
- Tried and trusted by researchers since 2003
IgG
Rabbit
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA
Liquid
Polyclonal
ICC/IF | WB | IHC-P | |
---|---|---|---|
Human | Tested | Tested | Tested |
Mouse | Expected | Tested | Expected |
Rat | Expected | Tested | Expected |
Hamster | Predicted | Predicted | Predicted |
Pig | Predicted | Predicted | Predicted |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 0.5-1 µg/mL | Notes Methanol fixation recommended. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Hamster, Pig | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/1000 - 1/2000 | Notes - |
Species Rat | Dilution info 1/1000 - 1/2000 | Notes - |
Species Human | Dilution info 1/1000 - 1/2000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Hamster, Pig | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/200 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Hamster, Pig | Dilution info - | Notes - |
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Serine/threonine-protein kinase component of the chromosomal passenger complex (CPC), a complex that acts as a key regulator of mitosis (PubMed:11516652, PubMed:12925766, PubMed:14610074, PubMed:14722118, PubMed:29449677). 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 (PubMed:11516652, PubMed:12925766, PubMed:14610074, PubMed:14722118, PubMed:26829474). Involved in the bipolar attachment of spindle microtubules to kinetochores and is a key regulator for the onset of cytokinesis during mitosis (PubMed:15249581). Required for central/midzone spindle assembly and cleavage furrow formation (PubMed:12458200, PubMed:12686604). Key component of the cytokinesis checkpoint, a process required to delay abscission to prevent both premature resolution of intercellular chromosome bridges and accumulation of DNA damage: phosphorylates CHMP4C, leading to retain abscission-competent VPS4 (VPS4A and/or VPS4B) at the midbody ring until abscission checkpoint signaling is terminated at late cytokinesis (PubMed:22422861, PubMed:24814515). AURKB phosphorylates the CPC complex subunits BIRC5/survivin, CDCA8/borealin and INCENP (PubMed:11516652, PubMed:12925766, PubMed:14610074). Phosphorylation of INCENP leads to increased AURKB activity (PubMed:11516652, PubMed:12925766, PubMed:14610074). Other known AURKB substrates involved in centromeric functions and mitosis are CENPA, DES/desmin, GPAF, KIF2C, NSUN2, RACGAP1, SEPTIN1, VIM/vimentin, HASPIN, and histone H3 (PubMed:11756469, PubMed:11784863, PubMed:11856369, PubMed:12689593, PubMed:14602875, PubMed:16103226, PubMed:21658950). A positive feedback loop involving HASPIN and AURKB contributes to localization of CPC to centromeres (PubMed:21658950). Phosphorylation of VIM controls vimentin filament segregation in cytokinetic process, whereas histone H3 is phosphorylated at 'Ser-10' and 'Ser-28' during mitosis (H3S10ph and H3S28ph, respectively) (PubMed:11784863, PubMed:11856369). AURKB is also required for kinetochore localization of BUB1 and SGO1 (PubMed:15020684, PubMed:17617734). Phosphorylation of p53/TP53 negatively regulates its transcriptional activity (PubMed:20959462). Key regulator of active promoters in resting B- and T-lymphocytes: acts by mediating phosphorylation of H3S28ph at active promoters in resting B-cells, inhibiting RNF2/RING1B-mediated ubiquitination of histone H2A and enhancing binding and activity of the USP16 deubiquitinase at transcribed genes (By similarity). Acts as an inhibitor of CGAS during mitosis: catalyzes phosphorylation of the N-terminus of CGAS during the G2-M transition, blocking CGAS liquid phase separation and activation, and thereby preventing CGAS-induced autoimmunity (PubMed:33542149). Phosphorylates KRT5 during anaphase and telophase (By similarity). Phosphorylates ATXN10 which promotes phosphorylation of ATXN10 by PLK1 and may play a role in the regulation of cytokinesis and stimulating the proteasomal degradation of ATXN10 (PubMed:25666058).
AIK2, AIM1, AIRK2, ARK2, STK1, STK12, STK5, AURKB, Aurora kinase B, Aurora 1, Aurora- and IPL1-like midbody-associated protein 1, Aurora/IPL1-related kinase 2, STK-1, Serine/threonine-protein kinase 12, Serine/threonine-protein kinase 5, Serine/threonine-protein kinase aurora-B, AIM-1, ARK-2, Aurora-related kinase 2
Anti-Aurora B antibody ab2254 is a rabbit polyclonal antibody that is used in Aurora B western blotting, IHC and immunofluorescence. Suitable for human, mouse and rat samples.
- Tried and trusted by researchers since 2003
IgG
Rabbit
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA
Liquid
Polyclonal
Affinity purification Immunogen
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
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This supplementary information is collated from multiple sources and compiled automatically.
Aurora B also known as Aurora kinase B or AURKB is a serine/threonine protein kinase with a molecular mass of approximately 39 kDa. This protein expresses across various cell types and tissues playing a critical role during cell division. It localizes to the centromeres during early mitosis and later associates with the central spindle and midbody. Aurora B is a component of the chromosomal passenger complex (CPC) which includes other proteins such as INCENP survivin and borealin essential for its function.
Aurora B kinase functions to ensure proper chromosome alignment segregation and cytokinesis during mitosis and meiosis. It phosphorylates various substrates to facilitate processes like correction of kinetochore-microtubule attachments and regulation of the mitotic checkpoint. As a part of the CPC Aurora B acts in coordination with other proteins to control these cellular events ensuring that cells divide accurately and safely.
Aurora B integrates into the cell cycle and mitotic pathways operating closely with the spindle assembly checkpoint. It interacts with proteins like CENP-A and CENP-E ensuring error-free chromosome segregation. Aurora B's activity within these pathways is critical for maintaining genomic stability and preventing aneuploidy a condition linked to improper chromosome number.
Aurora B's malfunction associates with cancer and chromosomal instability. Overexpression or dysregulation of Aurora B kinase frequently occurs in several types of cancer contributing to tumorigenesis by promoting abnormal cell division. Proteins such as p53 a well-known tumor suppressor often become functionally compromised when Aurora B activity is altered further advancing disease progression. Aurora B also holds implications in other disorders marked by faulty cell cycle regulation emphasizing its potential as a therapeutic target.
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AURKB is expressed in mouse oocytes.
(Panel D) 20 GV-intact oocytes were collected from CF1 mice and micro-injected with the indicated cRNA. Two hours after injection, the oocytes were matured to Met II in vitro (16 h). The total numbers of non-injected control oocytes (Non-inj.) are indicated in parenthesis. Total cellular lysates were probed with the indicated antibody. The panels are images of the same membrane that was stripped and re-probed. The arrows indicate the specific AURKB protein band, and the asterisk indicates a presumed degradation product of AURKB-GFP.
All lanes: Western blot - Anti-Aurora B antibody (ab2254)
Predicted band size: 39 kDa
AURKB is expressed in mouse oocytes.
(Panel A) GV-intact oocytes were collected from CF1 mice and matured in vitro for 8 h (Met I), or 16 h (Met II), prior to fixation and staining with an anti-AURKB antibody (ab2254).
ab2254 stained in Hela cells. Cells were fixed with 4% paraformaldehyde (10min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab2254 at 1μg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control) at 1/1000 dilution overnight at +4°C. The secondary antibodies were Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (pseudo-colored red) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 (colored green) used at 1 ug/ml for 1hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43μM for 1hour at room temperature.
HCT 116 (Human colorectal carcinoma cell line) cells were examined by immunofluorescence confocal microscopy.
Cells were treated with 500 nM AK301 for 16 hours, and then processed for Aurora B (ab2254) and β-tubulin staining (Panel B). The color key and 20 μm bars are shown. Images of representative field is shown with a 20 μm bar. End-labeled DNA is shown in red and DAPI-stained DNA is blue.
Cells cultured on coverslips were fixed with 4% paraformaldehyde at room temperature or 100% ice cold methanol at 4°C and then permeabilized with 0.5% Triton X-100 in PBS. Cells were blocked in 5% serum (in PBS) and then incubated with primary antibody (in 5% serum) on shaker for 1 h at room temperature.
Blocked with 2% BSA.
All lanes: Western blot - Anti-Aurora B antibody (ab2254) at 1 µg/mL
Lane 1: HeLa cell lysate at 10 µg
Lane 2: HeLa nocodozole treated cell lysate at 10 µg
Lane 3: NIH3T3 cell lysate at 10 µg
Lane 4: PC12 cell lysate at 10 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution
Performed under reducing conditions.
Predicted band size: 39 kDa
Immunofluorescence in human cells using Rabbit polyclonal to Aurora B (red), DAPI (blue) and CREST serum (binds to centromeres)(green).
(a) HeLa cells - transition from interphase (left) through mitosis
(b) RPE-1 cells - as in (a)
(c) HeLa cells - interphase
(d) RPE-1 cells - interphase
IHC image of Aurora B staining in Human Lymph node Hodgkins disease formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab2254, 5μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
ab2254 staining human A431 (epithelial) cells by ICC/IF. The sample was fixed in paraformaldehyde and permeabilized by incubation with 0.1% Triton X100. 1% BSA was used as the blocking agent prior to a 1 hour incubation with the primary antibody, diluted 1/1000 with 1% BSA made up in PBS. An Alexa Fluor® 647 conjugated Donkey anti-Rabbit IgG (H+L) antibody was used as the secondary. Blocking and antibody incubation steps were carried out at room temperature.
In this set of images, the tubulin is stained green, Aurora B in pink and DNA in blue.
Rabbit polyclonal to Aurora B (ab2254) used to stain SW620 human tumour xenografts (in mouse).
The sections were microwave pretreated in citrate buffer (pH 6.0) for 5 mins high then 5 mins simmer (800W conventional microwave). Slides were then incubated for 1 hour with the Aurora B primary antibody diluted 1/200 in TBS, then visualised using DAB, after application of an appropriate secondary.
All lanes: Western blot - Anti-Aurora B antibody (ab2254) at 1 µg/mL
Lane 1: HeLa Whole Cell Lysate at 20 µg
Lane 2: HeLa Nuclear Lysate at 20 µg
Lane 3: Jurkat Whole Cell Lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 39 kDa
Observed band size: 37 kDa, 39 kDa
Exposure time: 150s
All lanes: Western blot - Anti-Aurora B antibody (ab2254) at 1/2000 dilution
All lanes: Western blot - Recombinant human Aurora B protein (Recombinant human Aurora B protein ab51435) at 0.1 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (Goat Anti-Rabbit IgG H&L (HRP) preadsorbed ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 39 kDa
Exposure time: 30s
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