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Antibody Panel for the Detection of Aurora Kinase Family: Aurora A / STK15 / ARK1, Aurora B / STK12 / ARK2 and Aurora C / STK13 / AIK3

Chromosomal segregation during mitosis as well as meiosis is regulated by kinases and phosphatases. 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.

Zhou et al. (1998) found that STK15 is involved in the induction of centrosome duplication-distribution abnormalities and aneuploidy in mammalian cells. Centrosomes appear to maintain genomic stability through the establishment of bipolar spindles during cell division, ensuring equal segregation of replicated chromosomes to 2 daughter cells. Deregulated duplication and distribution of centrosomes are implicated in chromosome segregation abnormalities, leading to aneuploidy seen in many cancer cell types. In addition amplification of STK15 could be demonstrated in approximately 12% of primary breast tumors, as well as in breast, ovarian, colon, prostate, neuroblastoma, and cervical cancer cell lines. High expression of STK15 mRNA was detected in tumor cell lines without evidence of gene amplification. Ectopic expression of STK15 in mouse NIH 3T3 cells led to the appearance of abnormal centrosome number (amplification) and transformation in vitro. Finally, overexpression of STK15 in near-diploid human breast epithelial cells revealed similar centrosome abnormality, as well as induction of aneuploidy. Aurora kinase A is a key regulatory component of the p53 pathway and overexpression of aurora kinase A leads to increased degradation of p53, causing downregulation of checkpoint-response pathways and facilitating oncogenic transformation of cells. These findings suggested that STK15 is a critical kinase-encoding gene, whose overexpression leads to centrosome amplification, chromosomal instability, and transformation in mammalian cells.

Aurora Kinase B is a Ser/Thr protein kinase member of the Aurora subfamily that may be directly involved in regulating the cleavage of polar spindle microtubules and is a key regulator for the onset of cytokinesis during mitosis. Aurora Kinase B is localized to the midzone of central spindle in late anaphase and concentrated into the midbody in telophase and cytokinesis and is colocalized with gamma tubulin in the mid-body. High levels of Aurora B expression are seen in the thymus, although it is also expressed in the spleen, lung, testis, colon, placenta and fetal liver. Aurora B is expressed during S and G2/M phase and expression is up-regulated in cancer cells during M phase.

Figure. Figure shows a model of Aurora B, INCENP, and Survivin interactions and regulation. Survivin binds the N-terminus of INCENP, whereas the N terminus of Aurora B binds the C-terminus of INCENP
Figure shows a model of Aurora B, INCENP, and Survivin interactions and regulation. Survivin binds the N-terminus of INCENP, whereas the N terminus of Aurora B binds the C-terminus of INCENP

Human Aurora Kinase C (AURKC, STK13) acts as a chromosomal passenger protein and forms complexes with AURKB and INCENP. AURKC phosphorylated histone H3 in vitro. INCENP bound and activated AURKC in vivo and in vitro, and AURKC coexpressed with INCENP elicited histone H3 phosphorylation of interphase cells. Using an in vitro kinase assay, Yan et al. (2005) found that AURKC-SV could phosphorylate a test substrate protein, and its activity depended on thr179. During cell mitosis, AURKC-SV associated with chromosomes in prophase and metaphase, and then transferred to the central spindle midzone and the cortex where the contracting ring formed during the transition from anaphase to telophase. It remained in the midbody during cytokinesis.

Antibody Panel to Aurora Kinase Subfamily (Aurora A / STK15 antibody, Aurora B / STK12 antibody, Aurora C / STK13 antibody)

Acris Antibodies offers a collection of polyclonal and monoclonal antibodies to members of the Aurora kinase family (Aurora A / STK15 antibody, Aurora B / STK12 antibody, Aurora C / STK13 antibody) . These antibodies are usefull for detection of Aurora Kinases using western blotting and immunohistochemistry methods, but also other immunological applications, like immunocytochemistry and immunoprecipitation.



Panel A. Extracts from untreated Hela cells (Lane 1) or overnight nocodazole-treated Hela cells (Lane 2) were immunoprecipitated with a pan-Aurora A monoclonal antibody (clone 35C1), resolved by electrophoresis, transferred to nitrocellulose and probed with rabbit polyclonal antibody against Thr288 phosphorylated Aurora A. Proteins were visualized using a donkey anti-rabbit secondary conjugated to HRP and a chemiluminescence detection system.

Panel A. Extracts from untreated Hela cells (Lane 1) or overnight nocodazole-treated Hela cells (Lane 2) were immunoprecipitated with a pan-Aurora A monoclonal antibody (clone 35C1, NB100-1641), resolved by electrophoresis, transferred to nitrocellulose and probed with rabbit polyclonal antibody against Thr288 phosphorylated Aurora A NB600-1157. Proteins were visualized using a donkey anti-rabbit secondary conjugated to HRP and a chemiluminescence detection system.

Panel B. The blot shown in Panel A was stripped and re-probed with pan-Aurora specific polyclonal antibody to verify equal loading of Aurora A in the extracts. Proteins were visualized using a donkey anti-rabbit secondary conjugated to HRP and a chemiluminescence detection system.

Panel B. The blot shown in Panel A was stripped and re-probed with pan-Aurora specific polyclonal antibody to verify equal loading of Aurora A in the extracts. Proteins were visualized using a donkey anti-rabbit secondary conjugated to HRP and a chemiluminescence detection system.

<b>Detection of Human Phospho-Aurora A (T288) by  Immunocytochemistry.</b> <i>Samples:</i> NBF-fixed asynchronous  HeLa cells. <i>Antibody:</i> Affinity purified rabbit anti-Phospho-  Aurora A (T288) (NB 100-2371) used at a dilution of  1:250. <i>Detection:</I> Anti-rabbit IgG-FITC conjugated (NB 7159) used at a dilution of 1:100.

Detection of Human Phospho-Aurora A (T288) by Immunocytochemistry. Samples: NBF-fixed asynchronous HeLa cells. Antibody: Affinity purified rabbit anti-Phospho- Aurora A (T288) (NB 100-2371) used at a dilution of 1:250. Detection: Anti-rabbit IgG-FITC conjugated (NB 7159) used at a dilution of 1:100.

<b>Detection of Human Phospho-Aurora A (T288) by  Immunocytochemistry.</b> <i>Samples:</i> NBF-fixed asynchronous  HeLa cells. Mock phosphatase treated section (left) or calf  intestinal phosphatase-treated section (right) immunostained for  Phospho-Aurora A. <i>Antibody:</i> Affinity purified rabbit anti-  Phospho-Aurora A (T288) (NB 100-2371) used at a  dilution of 1:250. <i>Detection:</i> Anti-rabbit IgG-FITC conjugated  (NB 7159) used at a dilution of 1:100.

Detection of Human Phospho-Aurora A (T288) by Immunocytochemistry. Samples: NBF-fixed asynchronous HeLa cells. Mock phosphatase treated section (left) or calf intestinal phosphatase-treated section (right) immunostained for Phospho-Aurora A. Antibody: Affinity purified rabbit anti- Phospho-Aurora A (T288) (NB 100-2371) used at a dilution of 1:250. Detection: Anti-rabbit IgG-FITC conjugated (NB 7159) used at a dilution of 1:100.

Detection of rat Aurora A in mammary gland whole cell tissue lysate (60 ug) using NB 100-635.  6 minute ECL exposure time. *Note: The upper band is believed to be phosphorylated Aurora A.

Detection of rat Aurora A in mammary gland whole cell tissue lysate (60 ug) using NB 100-635. 6 minute ECL exposure time. *Note: The upper band is believed to be phosphorylated Aurora A.

Immunofluorescent localization of mouse Aurora A to centrosomes of a mammary epithelial cell line using NB 100-635.  Detection by Alexa 488 labeled gaot anti-Rabbit IgG (green) and DAPI (blue).

Immunofluorescent localization of mouse Aurora A to centrosomes of a mammary epithelial cell line using NB 100-635. Detection by Alexa 488 labeled gaot anti-Rabbit IgG (green) and DAPI (blue).

<b>Detection of Human Aurora B by Immunohistochemistry.</b> <i>Sample:</i> FFPE section of human colon adenocarcinoma. <i>Antibody:</i> Affinity purified rabbit anti-Aurora B NB 100-1729 used at a dilution of 1:100. <i>Detection:</i> DAB

Detection of Human Aurora B by Immunohistochemistry. Sample: FFPE section of human colon adenocarcinoma. Antibody: Affinity purified rabbit anti-Aurora B NB 100-1729 used at a dilution of 1:100. Detection: DAB



Available antibodies to Aurora Kinases: Aurora A antibody, STK15 antibody, ARK1 antibody, Aurora B antibody, STK12 antibody, ARK2 antibody, Aurora C antibody, STK13 antibody, AIK3 antibody



 

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Antibodies


Catalog No. Clone/Isotype Presentation Reactivity Applications  
SM2084B Aurora-A / STK15 / ARK1  
  35C1 Biotin Hu, Ms IF, WB 0.1 mg / 390 €
Mouse IgG2b Acris Antibodies GmbH
 
SM2084P Aurora-A / STK15 / ARK1  
  35C1 Purified Hu, Ms IF, IP, WB 0.1 mg / 360 €
Mouse IgG2b Acris Antibodies GmbH
 
SP2058P Aurora-A / STK15 / ARK1  
  - Aff - Purified Hu C, IP 50 µg / 280 €
Rabbit Acris Antibodies GmbH
 
EB05035 Aurora-A / STK15 / ARK1  
  - Aff - Purified Hu WB 0.1 mg / 300 €
Goat Everest Biotech Ltd.
 
NB100-1641 Aurora-A / STK15 / ARK1  
  35C1 Purified Hu, Ms E, IF, IP, WB 50 µl / 330 €
Mouse IgG2b Novus Biologicals Inc.
 
NB100-1829 Aurora-A / STK15 / ARK1  
  35C1 Biotin Hu, Ms IF, WB 50 µl / 360 €
Mouse IgG2b Novus Biologicals Inc.
 
NB100-212 Aurora-A / STK15 / ARK1  
  - Aff - Purified Hu C, IP 0.1 ml / 360 €
Rabbit Novus Biologicals Inc.
 
NB100-267 Aurora-A / STK15 / ARK1  
  - Aff - Purified Hu IF, IP, WB 0.1 mg / 360 €
Rabbit Novus Biologicals Inc.
 
NB100-635 Aurora-A / STK15 / ARK1  
  - Aff - Purified Ms, Rt IF, WB 0.1 mg / 330 €
Rabbit Novus Biologicals Inc.
 
NB100-917 Aurora-A / STK15 / ARK1  
  Aff - Purified Hu, Ms, Rt WB, IP, IF 3 x 0.1 ml / 610 €
Rabbit Novus Biologicals Inc.
 
NB600-435 Aurora-A / STK15 / ARK1  
  - Agarose Hu IP 0.4 ml / 380 €
Rabbit Novus Biologicals Inc.
 
10297-1-AP Aurora-A / STK15 / ARK1  
  Aff - Purified Hu, Ms E, P, WB 150 µl / 320 €
Rabbit IgG ProteinTech Group, Inc.
 
NB110-55479 Aurora-A / STK15 / ARK1 (C-term)  
  EP1008Y Purified Hu IP, WB, IF 0.1 ml / 410 €
Rabbit IgG Novus Biologicals Inc.
 
13040 Aurora-A / STK15 / ARK1 (exon 5)  
  - Aff - Purified Hu C, IP, WB 0.1 mg / 340 €
Rabbit QED Bioscience Inc.
 
13042 Aurora-A / STK15 / ARK1 (exons 5, 6)  
  - Aff - Purified Hu IF, IP, WB 0.1 mg / 340 €
Rabbit QED Bioscience Inc.
 
NB100-2371 Aurora-A / STK15 / ARK1 (pThr288)  
  - Purified Hu IF 0.1 ml / 410 €
Rabbit Novus Biologicals Inc.
 
NB600-1157 Aurora-A / STK15 / ARK1 (pThr288)  
  - Purified Hu WB 0.1 ml / 380 €
Rabbit Novus Biologicals Inc.
 
SP7343P Aurora-A/B / STK12/15 / ARK1/2  
  - Purified Hu, Ms, Rt WB 0.1 mg / 280 €
Rabbit Acris Antibodies GmbH
 
BP4524 Aurora-B / STK12 / ARK2  
  - Aff - Purified Hu WB 0.2 ml / 380 €
Sheep Acris Antibodies GmbH
 
R1539P Aurora-B / STK12 / ARK2  
  - Aff - Purified Hu E, WB 0.1 mg / 340 €
Rabbit Acris Antibodies GmbH
 
NB100-1729 Aurora-B / STK12 / ARK2  
  Aff - Purified Hu P 0.1 ml / 410 €
Rabbit Novus Biologicals Inc.
 
NB100-294 Aurora-B / STK12 / ARK2  
  - Purified Hu, Ms, Rt, Hst IF, IP, WB 0.1 ml / 380 €
Rabbit Novus Biologicals Inc.
 
NB500-185 Aurora-B / STK12 / ARK2  
  - Aff - Purified Hu IF, IP, WB 0.1 mg / 330 €
Rabbit Novus Biologicals Inc.
 
10792-1-AP Aurora-B / STK12 / ARK2  
  Aff - Purified Hu, Ms E 150 µl / 320 €
Rabbit IgG ProteinTech Group, Inc.
 
65031-1-Ig Aurora-B / STK12 / ARK2  
  Purified Hu E, WB 150 µl / 320 €
Mouse IgG1 ProteinTech Group, Inc.
 
NB110-55480 Aurora-B / STK12 / ARK2 (N-term)  
  EP1009Y Purified Hu IP, WB, F, IF 0.1 ml / 380 €
Rabbit IgG Novus Biologicals Inc.
 
A4190-17 Aurora-C / STK13 / AIK3  
  - Aff - Purified Hu E 40 µg / 220 €
Rabbit United States Biological
 
NB110-55481 Aurora-C / STK13 / AIK3 (N-Term)  
  EP1011Y Purified Hu WB 0.1 ml / 380 €
Rabbit IgG Novus Biologicals Inc.