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>   home   >   Products   >   Primary Antibodies   >   Signal Transduction   >   Anti Aurora-A Kinase Antibody, clone 35C1    

Anti Aurora-A Kinase Antibody, clone 35C1

Mouse Anti-Human Monoclonal Antibody

     
  • WB - Anti Aurora-A Kinase Antibody, clone 35C1  ABD13399
    Recombinant Aurora kinase detected with Mouse anti Aurora-A kinase
  • IF - Anti Aurora-A Kinase Antibody, clone 35C1  ABD13399
    Published customer image:Mouse anri Human Aurora -A kinase monoclonal antibody, clone 35C1 used for the evaluation of Aurora A kinase expression in HeLa cells during mitosis by immunofluorescence microscopy, immunoprecipitation and western blotting.Image caption:Aurora A localizes with and phosphorylates centrin in vitro and in cells. (A) Immunofluorescence confocal microscopy of HeLa cells demonstrates that Aurora A and phospho-S170 centrin both localize at mitotic spindle poles from prophase through metaphase. Phospho-centrin is greatly reduced in anaphase and telophase and Aurora A is greatly reduced by telophase. DNA was counterstained with DAPI . Scale bar?=?10 microns. (B) Western blots of whole cell lysates from cycling and nocodazole-arrested mitotic cells show that centrin and Aurora A are present in asynchronous and synchronous cultures, with greater amounts of Aurora A in mitotic cells. Reciprocal immunoprecipitations of mitotic cell lysates demonstrate an interaction between Aurora A and centrin during mitosis. (C) In in vitro kinase assays with recombinant centrin incubated with recombinant Aurora A, centrin was phosphorylated only in the presence of ATP. Under conditions containing ATP, a shift in centrin is detected when Western blotted with a total centrin antibody (lower panel) and phospho-centrin is detected by the phospho-S170 centrin antibody (upper panel). Phospho-centrin is not detected under conditions lacking Aurora A and/or ATP. (D) In vitro kinase assays using endogenous Aurora A immunoprecipitated from nocodazole-arrested HeLa cells generated phospho-centrin only in the presence of ATP as demonstrated by Western blotting with the phospho-S170 centrin antibody. (E) Kinase-active Aurora A is required for phosphorylation of centrin at mitotic spindle poles. Cells that lack endogenous Aurora A but express shRNA-resistant GFP-WT Aurora A clearly exhibit phospho-S170 centrin staining; while those that lack endogenous Aurora A and express GFP-kinase dead Aurora A exhibit a nearly complete loss of phospho-S170 centrin staining at the mitotic spindle poles. White arrows denote the focal staining of gamma-tubulin at poles in the cell expressing WT Aurora A or kinase dead Aurora A . From: Lukasiewicz KB, Greenwood TM, Negron VC, Bruzek AK, Salisbury JL, Lingle WL Control of Centrin Stability by Aurora A.PLoS ONE 6(6): e21291.
  • WB - Anti Aurora-A Kinase Antibody, clone 35C1  ABD13399
    Published customer image:Mouse anri Human Aurora -A kinase monoclonal antibody, clone 35C1 used for the evaluation of Aurora A kinase expression in HeLa cells during mitosis by western blotting.Image caption:Centrin interacts with APC/C during mitosis. (A) Levels of Aurora A, centrin, and cyclin B were compared in Western blots of lysates of HeLa cells harvested at the indicated time points after synchronization by double thymidine/nocodazole block and release. Cyclin B degradation is used to indicate the onset of anaphase, while the beta-actin blot serves as a loading control. Aurora A and centrin levels drop to basal levels by 150 minutes post-release, while cyclin B is at basal levels by 120 minutes post-release. (B) Equal volumes of immunoprecipitations of lysates from cycling (C) or nocodazole arrested (N) HeLa cells performed with Cdc20 and centrin and Western blotted with the indicated antibodies demonstrate that centrin is pulled down with cdc20 only in nocodazole arrested cells along with Cdc16 and Cdc27. Cdc20 is pulled down with centrin in nocodazole arrested cells and to a slightly lesser extent in cycling cells. The antibody heavy chain and cdc20 were indicated with (grey triangle) and (dark triangle), respectively. (C) In lysates from asynchronously growing HeLa cells only non-phosphorylated centrin immunoprecipitates with cdc20. Cdc20 does not pull down phosphorylated centrin even though p-S170 centrin is abundant, as seen in the lysate-only lane. (D) HeLa Tet-On cells expressing various centrin mutants treated overnight with DMSO (D), ALLnL (A), leupeptin (L), MG132 (M), and ammonium chloride (N) and Western blotted with antibodies directed against total centrin reveal that DMSO, leupeptin, and ammonium chloride do not prevent centrin degradation, whereas ALLnL and MG132, the two proteasome inhibitors, do prevent degradation of wildtype and mutant forms of centrin. Lamin B loading controls are shown for each mutant cell line. (E) Lysates from HeLa Tet-On cells expressing S170A centrin treated with DMSO (D) or ALLnL (A) for 16 hours were Western blotted for centrin and beta-actin show significant degradation products in the presence of ALLnL but not DMSO. Additionally, when the boxed area of the centrin blot is over-exposed, 7 kDa laddering indicative of ubiquitination is evident (open triangle). Endogenous and HA-centrin are indicated with (light triangle) and (dark triangle), respectively.From: Lukasiewicz KB, Greenwood TM, Negron VC, Bruzek AK, Salisbury JL, Lingle WL Control of Centrin Stability by Aurora A.PLoS ONE 6(6): e21291.
  • WB - Anti Aurora-A Kinase Antibody, clone 35C1  ABD13399
    Published customer image:Mouse anri Human Aurora -A kinase monoclonal antibody, clone 35C1 used for the evaluation of Aurora A kinase expression in HeLa cells during mitosis by western blotting.Image caption:Aurora A knock down in HeLa cells. HeLa cells were transfected with Aurora A shRNA and whole cell lysates were harvested 24 hours after transfection. Whole cell lysates were separated by SDS-PAGE and blotted with the indicated antibodies.From: Lukasiewicz KB, Greenwood TM, Negron VC, Bruzek AK, Salisbury JL, Lingle WL Control of Centrin Stability by Aurora A.PLoS ONE 6(6): e21291.
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Product Information
Application
  • Applications Legend:
  • WB=Western Blot
  • IHC=Immunohistochemistry
  • IHC-P=Immunohistochemistry (Paraffin-embedded Sections)
  • IHC-F=Immunohistochemistry (Frozen Sections)
  • IF=Immunofluorescence
  • FC=Flow Cytopmetry
  • IC=Immunochemistry
  • ICC=Immunocytochemistry
  • E=ELISA
  • IP=Immunoprecipitation
  • DB=Dot Blot
  • CHIP=Chromatin Immunoprecipitation
  • FA=Fluorescence Assay
  • IEM=Immunoelectronmicroscopy
  • EIA=Enzyme Immunoassay
WB, IF, IP
Primary Accession O14965
Reactivity Human
Host Mouse
Clonality Monoclonal
Isotype IgG2b
Clone Names 35C1
Calculated MW 45809 Da
Additional Information
Other Species M
Purification Purified IgG prepared by affinity chromatography on Protein G
Immunogen Recombinant Aurora-A
Shelf Life 18 months from date of despatch.
Gene ID 6790
Other Names Aurora kinase A, 2.7.11.1, Aurora 2, Aurora/IPL1-related kinase 1, ARK-1, Aurora-related kinase 1, hARK1, Breast tumor-amplified kinase, Serine/threonine-protein kinase 15, Serine/threonine-protein kinase 6, Serine/threonine-protein kinase aurora-A, AURKA
Target/Specificity Mouse anri Human Aurora -A kinase monoclonal antibody, clone 35C1 recognizes human Aurora-A kinase also known as Aurora 2, breast tumor-amplified kinase and serine/threonine-protein kinase 6 or 15. Aurora kinase A is member of the Ser/Thr protein kinase family containing a single protein kinase domain, has a molecular weight of ~46kDa and is involved in mitotic spindle assembly (Ducatet al.2004).Aurora-A kinase is reported to be overexpressed in many epithelial cancers and is thought to play an important role in tumourigenesis (Katayamaet al.2003). Aurora A kinase appears to facilitate phosphorylation of centrin and co-localizes with it at centrosomes with maximum expression through prophase to late metaphase (Lukasiewiczet al.2011).Mouse anti human Aurora-A kinase, clone 35C1 specifically recognizes an epitope within the non-catalytic N-terminal domain of Aurora-A. Clone 35C1 does not inhibit Aurora-A kinase activity (Cremetet al.2003).
Preservative & Stabilisers 0.09% Sodium Azide (NaN3)
Storage Store at +4℃ or at -20℃ if preferred.
PrecautionsAnti Aurora-A Kinase Antibody, clone 35C1 is for research use only and not for use in diagnostic or therapeutic procedures.
Research Areas
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