Aurora Kinase B (AURKB) (pThr232) antibody

Antigen: Aurora Kinase B (AURKB)

Synonyms: AIK2, AIM1, ARK2, AurB, IPL1, STK5, AIM-1, STK12, aurkb-sv1, aurkb-sv2, AIE1, AIK3, Stk13, Aik2, Aim1, Ark2, Stk5, AIRK2, STK-1, Stk12, AL022959, DmAIRK2, DmAurB, DmAuroraB, IAL, IAL1, aurora B, DmelCG6620, CG6620, AURKB, MGC142782, aik2, aim1, ark2, ipl1, stk5, aim-1, stk12, Aurora-B, fa09g06, MGC85918, CHUNP6896, wu:fa09g06, aurkb

Epitope: pThr232

Clonality: Polyclonal

Host: Rabbit

Reactivity: Cow (Bovine)

Application: Enzyme Immunoassay (EIA), Western Blotting (WB)

Catalog no.: ABIN401272

Additional Information

Characteristics: Synonyms: Aurora-B, Aurora/IPL1-related kinase 2, STK-1, AURKB, AIK2, AIM1, ARK2, STK12, Aurora- andIpl1-like midbody-associated protein 1, Serine/threonine-protein kinase 12

Alternative name: Aurora kinase B

Swiss-Prot: Q96GD4

Immunogen: Synthetic peptide corresponding aa 223-234 of Human Aurora Kinase B proteinAA Sequence: 1 maqkensypw pygrqtapsg lstlpqrvlr kepvtpsalv lmsrsnvqpt aapgqkvmen61 ssgtpdiltr hftiddfeig rplgkgkfgn vylarekksh fivalkvlfk sqiekegveh121 qlrreieiqa hlhhpnilrl ynyfydrrri ylileyaprg elykelqksc tfdeqrtati181 meeladalmy chgkkvihrd ikpenlllgl kgelkiadfg wsvhapslrr ktmcgtldyl241 ppemiegrmh nekvdlwcig vlcyellvgn ppfesashne tyrrivkvdl kfpasvptga301 qdliskllrh npserlplaq vsahpwvran srrvlppsal qsvaRemarks: 344 aa, predicted MW 39. 3kDa

Cross-Reactivity: Cow (Bovine), Dog (Canine), Chimpanzee, Human, Mouse (Murine), Pig (Porcine), Rat (Rattus)

Isotype: IgG  (Matching secondary antibodies)

Description: Aurora Kinase B (also known as Aurora and Ipl1-like midbody-associated protein 1, AIM-1,Aurora/IPL1-related kinase 2, Aurora-related kinase 2, STK-1, and Aurora-B) is a Ser/Thrprotein kinase member of the Aurora subfamily that may be directly involved in regulatingthe cleavage of polar spindle microtubules and is a key regulator for the onset ofcytokinesis during mitosis. Aurora Kinase B is localized to the midzone of central spindlein late anaphase and concentrated into the midbody in telophase and cytokinesis and iscolocalized with gamma tubulin in the mid-body. High levels of Aurora B expression areseen 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 isup-regulated in cancer cells during M phase.

Specificity: The product was affinity purified from monospecific antiserum by Immunoaffinitypurification. Antiserum was first purified against the phosphorylated form of theimmunizing peptide. The resultant affinity purified antibody was then cross-adsorbedagainst the non-phosphorylated form of the immunizing peptide. This antibody is directed against the phosphorylated form of human Aurora Kinase B at thepT232 residue. Reactivity with nonphosphorylated human Aurora Kinase B is minimal byELISA. No reaction is expected against Aurora Kinase A. Species Reactivity: Tested: Human. Expected from sequence similarity: Mouse, Rat, Bovine, Pig, Dog and Chimpanzee (100%). Add. Information: 1. Grow up 3X 10cm plates of MCF-7 cells in regular DMEM supplemented with 10% FBS. 2. Wash cells 2X with 1X PBS. 3. Lyse cells by adding 400 μl of Mammalian Protein Extraction Reagent (M-PER(TM),PIERCE) or PTY buffer (50mM HEPES pH7. 5, 50mM NaCl, 5mM EDTA, 1% Triton X-100, 50mMNaF, 10mM Na4P2O7 supplemented with protease inhibitors 1mM PMSF, 0. 01mg/mlleupeptine, 0. 01mg/ml aprotinin and phosphatase inhibitor 1mM Na3VO4). Let stand for5-10 min. 4. Scrape the cells using a Cell Lifter (Fisher). Transfer the lysate to a 1. 5 ml tube andcentrifuge at maximum speed (~14,000rpm) for 10 min at +4°C to clear the lysate. From thispoint on, protein lysate must be maintained on ice. 5. Transfer the supernatant (lysate) to a clean centrifuge tube. Perform a proteindetermination. Aliquot the lysate in 100 μg units. 6. Use 50-100 μg for the detection of total level of Aurora B protein or 200-500 μg forphosphorylated form (pT232) Aurora B. 7. Add 10 μl of 10X SDS-loading buffer and boil for 3 to 5 min in a water bath. 8. Load the content of the tube onto a 10% SDS-PAGE gel and separate by electrophoresis. 9. Transfer to the PVDF membrane (Millipore) in Tris-Glycine Buffer with 15% Methanol. DONOT ADD SDS! 10. Wash 2X in dH2O. 11. Incubate with blocking solution (5% BSA, 0. 1%Tween-20 in 1X TBS) for 1h. 12. Add primary Antibody for 1h. For total Aurora B use code #R1539P at a 1: 1,000 dilution. For phospho T232 detection use code #ABIN401272 at a 1: 250 dilution. Gently rockantibody-membrane reaction +4° C overnight. 13. Wash 3X in 1X TBS with 0. 1%Tween-20 for 10-15 min per wash. 14. Add secondary antibody HRP-Gt-a-RABBIT IgG (code #R1454HRP) at a 1: 10,000 dilutionfor 1h. 15. Wash 3X in 1XTBS with 0. 1%Tween-20. 16. Develop by ECL-Plus (Amersham) or West-Pico (PIERCE).

Synonyms: AIK2, AIM1, ARK2, AurB, IPL1, STK5, AIM-1, STK12, aurkb-sv1, aurkb-sv2, AIE1, AIK3, Stk13, Aik2, Aim1, Ark2, Stk5, AIRK2, STK-1, Stk12, AL022959, DmAIRK2, DmAurB, DmAuroraB, IAL, IAL1, aurora B, DmelCG6620, CG6620, AURKB, MGC142782, aik2, aim1, ark2, ipl1, stk5, aim-1, stk12, Aurora-B, fa09g06, MGC85918, CHUNP6896, wu:fa09g06, aurkb

Application Details

Application Notes: ELISA: 1/10,000-1/50,000. Western blot: 1/250-1/2,000 (See protocol below). Expect a band ~39 kDa in size corresponding to Aurora Kinase B by western blotting in theappropriate cell lysate or extract. HeLa cell lysate can be used as a positive control. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.

Concentration: 1.21 mg/ml (by UV absorbance at 280 nm)

Purification: Aff - Purified

Buffer: 0.02M Potassium Phosphate, 0.15M Sodium Chloride, pH 7.2, 0.01% (w/v)Sodium Azide as preservative.

Storage: Store the antibody undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer. Avoid repeated freezing and thawing. Should this product contain a precipitate werecommend microcentrifugation before use. Shelf life: one year from despatch.

Restrictions: For Research Use only