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Aurora-B Recombinant Rabbit mAb (SDT-079-32)

Aurora-B Recombinant Rabbit mAb (SDT-079-32)

Catalog Number: S0B2034 Application: WB,IHC-P,ICC,FCM,IP Reactivity: Human Conjugation: Unconjugated Brand: Starter
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Regular price $100.00 USD
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Product Details

Product Specification


Host Rabbit
Antigen Aurora-B
Synonyms Aurora 1, AIM-1, ARK-2, STK-1, Aurora-related kinase 2
Immunogen Synthetic Peptide
Accession Q96GD4
Clone Number SDT-079-32
Antibody Type Rabbit mAb
Application WB, IHC-P, ICC, ICFCM, IP
Reactivity Hu
Purification Protein A
Concentration 0.5mg/ml
Conjugation Unconjugated
Physical Appearance Liquid
Storage Buffer PBS, 40% Glycerol, 0.05%BSA, 0.03% Proclin 300
Stability & Storage 12 months from date of receipt / reconstitution, -20 °C as supplied

Dilution


application dilution species
IP 1:25
IHC-P 1:500-2000
WB 1:1000
ICC 1:500
ICFCM 1:5000

Background

Aurora kinase B is a protein that functions in the attachment of the mitotic spindle to the centromere. The expression and activity of Aurora B are regulated according to the cell cycle. Expression of Aurora B reaches a maximum at the G2-M transition, whereas Aurora B protein is most active during mitosis. The Aurora kinases associate with microtubules during chromosome movement and segregation. Aurora kinase B localizes to microtubules near kinetochores, specifically to the specialized microtubules called K-fibers, and Aurora kinase A localizes to centrosomes. In cancerous cells, over-expression of these enzymes causes unequal distribution of genetic information, creating aneuploid cells, a hallmark of cancer.

Picture

Western Blot

WB result of Aurora-B Rabbit mAb                

Primary antibody : Aurora-B Rabbit mAb at 1/1000 dilution
Lane 1: Hela whole cell lysate 20 µg
Lane 2: HepG2 whole cell lysate 20 µg
Lane 3: Raji whole cell lysate 20 µg
Lane 4: T47D whole cell lysate 20 µg
Lane 5: A549 whole cell lysate 20 µg

Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution

Predicted MW: 41 kDa
Observed MW: 39 kDa
Exposure time: 16 s

FC

Flow cytometric analysis of 4% PFA fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) labelling Aurora-B antibody at 1/5000 dilution (0.01 μg)/ (Right) compared with a Rabbit monoclonal IgG / (Left) isotype control. Goat Anti - Rabbit IgG Alexa Fluor® 488 was used as the secondary antibody. Cells were co-stained with DAPI to differentiate cell cycle phase.

IP

Aurora-B Rabbit mAb at 1/25 dilution (2µg) immunoprecipitating Aurora-B in 0.4mg HeLa whole cell lysate.
Western blot was performed on the immunoprecipitate using Aurora-B Rabbit mAb at 1/1000 dilution.
Secondary antibody (HRP) for IP was used at 1/400 dilution.
Lane 1 : HeLa whole cell lysate 10µg (input)
Lane 2 (+): Aurora-B Rabbit mAb IP in HeLa whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG IP in HeLa whole cell lysate
Predicted MW: 41 kDa
Observed MW: 39 kDa
Exposure time: 10s

Immunohistochemistry

IHC shows positive staining in paraffin-embedded human tonsil.

Anti-Aurora B antibody was used at 1/2000 dilution, followed by a Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Counterstained with hematoxylin.              

Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.

IHC shows positive staining in paraffin-embedded human cervix cancer.

Anti-Aurora B antibody was used at 1/500 dilution, followed by a Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Counterstained with hematoxylin.

Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.

Immunocytochemistry

ICC shows positive staining in HeLa cells. Anti-Aurora-B antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 4%PFA and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue).Counterstain with tubulin (red).

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