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Aurkb

Manufactured by Merck Group
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AURKB is a laboratory equipment product manufactured by Merck Group. It functions as a key regulator of cell division and is involved in the process of chromosome segregation during mitosis.

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Lab products found in correlation

Caliper labchip ez reader, by PerkinElmer (2 mentions) Phospho histone h3ser10, by Santa Cruz Biotechnology (1 mentions) Cleaved parp1 asp214, by BD (1 mentions) Aurka, by BD (1 mentions) Apoptosis western blot cocktail, by Abcam (1 mentions) β actin, by Merck Group (1 mentions) Cdc27, by BD (1 mentions) Survivin, by Merck Group (1 mentions)

Close spelling variants of this product

Anti-AURKB (2 citations)

4 protocols using aurkb

1

Enzyme Kinetic Assay for AURKB Inhibition

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Enzyme kinetic experiments were performed at pH 7.0 in 8 mM MOPS buffer with 0.2 mM EDTA and 10 mM magnesium acetate. Reactions were assembled in 384-well plate wells by adding 400 ng/ml of AURKB (EMD Millipore cat. no. 14-835) into separate reaction mixtures containing 1.5μM fluorescently labeled Caliper peptide substrate (FL-peptide 1, 5-FAM-AKRRRLSSLRA-COOH, Perkin-Elmer cat. no. 760345) with various concentrations of ATP and compound (BRD-7880, tozasertib, barasertib). The final ATP concentrations varied from 6.25 to 200 μM and compound varied from 0 to 200 nM. Plates were immediately placed into a Perkin-Elmer Caliper LabChip EZ Reader and wells were sampled periodically throughout a 1-hour reaction period for initial reaction rate. The fluorescent product and substrate were separated and monitored on the Caliper microfluidic instrument. The conversion of substrate was calculated with Caliper software. Km and ki values were determined from the double reciprocal Lineweaver-Burk plot by linear regression with GraFit 6 software (Erithacus Software Ltd., Horley, U.K.) using competitive inhibition equation modeling.
Yu C., Mannan A.M., Yvone G.M., Ross K.N., Zhang Y.L., Marton M.A., Taylor B.R., Crenshaw A., Gould J.Z., Tamayo P., Weir B.A., Tsherniak A., Wong B., Garraway L.A., Shamji A.F., Palmer M.A., Foley M.A., Winckler W., Schreiber S.L., Kung A.L, & Golub T.R. (2016). High-throughput identification of genotype-specific cancer vulnerabilities in mixtures of barcoded tumor cell lines. Nature biotechnology, 34(4), 419-423.
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2

Enzyme Kinetic Assay for AURKB Inhibition

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Enzyme kinetic experiments were performed at pH 7.0 in 8 mM MOPS buffer with 0.2 mM EDTA and 10 mM magnesium acetate. Reactions were assembled in 384-well plate wells by adding 400 ng/ml of AURKB (EMD Millipore cat. no. 14-835) into separate reaction mixtures containing 1.5μM fluorescently labeled Caliper peptide substrate (FL-peptide 1, 5-FAM-AKRRRLSSLRA-COOH, Perkin-Elmer cat. no. 760345) with various concentrations of ATP and compound (BRD-7880, tozasertib, barasertib). The final ATP concentrations varied from 6.25 to 200 μM and compound varied from 0 to 200 nM. Plates were immediately placed into a Perkin-Elmer Caliper LabChip EZ Reader and wells were sampled periodically throughout a 1-hour reaction period for initial reaction rate. The fluorescent product and substrate were separated and monitored on the Caliper microfluidic instrument. The conversion of substrate was calculated with Caliper software. Km and ki values were determined from the double reciprocal Lineweaver-Burk plot by linear regression with GraFit 6 software (Erithacus Software Ltd., Horley, U.K.) using competitive inhibition equation modeling.
Yu C., Mannan A.M., Yvone G.M., Ross K.N., Zhang Y.L., Marton M.A., Taylor B.R., Crenshaw A., Gould J.Z., Tamayo P., Weir B.A., Tsherniak A., Wong B., Garraway L.A., Shamji A.F., Palmer M.A., Foley M.A., Winckler W., Schreiber S.L., Kung A.L, & Golub T.R. (2016). High-throughput identification of genotype-specific cancer vulnerabilities in mixtures of barcoded tumor cell lines. Nature biotechnology, 34(4), 419-423.
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3

Western Blot Analysis of Cell Cycle Markers

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Antibodies against CDK1 [47 (link)] and cyclin B1 [41 (link)] were obtained from sources as described previously. Antibodies against β-actin (Sigma-Aldrich), AURKA, CDC27, cleaved PARP1(Asp214), and phospho-PLK1Thr210 (BD Biosciences, Franklin Lakes, NJ, USA), AURKB (Sigma-Aldrich), phospho-AURKAThr288/AURKBThr232/AURKCThr198 (Cell Signaling Technology, Beverly, MA, USA), cleaved caspase 3 (using a Apoptosis Western Blot Cocktail, Abcam, Cambridge, UK), phospho-histone H3Ser10 and PLK1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA) were obtained from the indicated suppliers. Immunoblotting was performed as described [44 (link)].
Li J., Hong M.J., Chow J.P., Man W.Y., Mak J.P., Ma H.T, & Poon R.Y. (2015). Co-inhibition of polo-like kinase 1 and Aurora kinases promotes mitotic catastrophe. Oncotarget, 6(11), 9327-9340.
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4

Lentiviral shRNA Knockdown Targets

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pLKO.1 lenti-viral vectors for shRNAs targeting mouse Cdk1, Aurkb, Incenp, Borealin and Survivin were purchased from Sigma-Aldrich MISSON® library. shRNA sequences used in this study are as followed. shCdk1 #1(TRCN0000274559), #3 (TRCN0000274503), shAurkb #2 (TRCN0000374361), #4 (TRCN0000321651), shINCENP #1 (TRCN0000072093), #2 (TRCN0000072094), shBorealin #1 (TRCN0000177470), #2 (TRCN0000177578), shSurvivin #3 (TRCN0000054617). For shSurvivin #18 (5′-CCGCATCTCTACATTCAAGAA-3′) (20 (link)), annealed oligomers were cloned into pLKO.1 vector. pLKO.1 puro vector was used as a control.
Kim H.J., Shin J., Lee S., Kim T.W., Jang H., Suh M.Y., Kim J.H., Hwang I.Y., Hwang D.S., Cho E.J, & Youn H.D. (2018). Cyclin-dependent kinase 1 activity coordinates the chromatin associated state of Oct4 during cell cycle in embryonic stem cells. Nucleic Acids Research, 46(13), 6544-6560.
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