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Biochemistry
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JoVE Journal Biochemistry
Detection of Small GTPase Prenylation and GTP Binding Using Membrane Fractionation and GTPase-linked Immunosorbent Assay
 

Detection of Small GTPase Prenylation and GTP Binding Using Membrane Fractionation and GTPase-linked Immunosorbent Assay

Article DOI: 10.3791/57646-v • 13:52 min • November 11th, 2018
November 11th, 2018 •
1,2, 1,2,3, 1, 1,2, 4, 5, 6,7,8, 1,9, 1,2,10
1Department of Human Anatomy and Cell Science, Rady Faculty of Health Sciences, Max Rady College of Medicine, University of Manitoba, 2Biology of Breathing Theme, Children Hospital Research Institute of Manitoba, University of Manitoba, 3Department of Biochemistry, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, 4Division of Pulmonary, Critical Care, and Sleep Medicine, Department of Internal Medicine, Center for Comparative Respiratory Biology and Medicine, 5Department of Clinical Biochemistry, School of Medicine, Zahedan University of Medical Sciences, 6Department of Molecular Biology, School of Pharmacy with the Division of Laboratory Medicine in Sosnowiec, Medical University of Silesia, 7Centre de biophysique moléculaire - UPR 4301, Centre national de la recherche scientifique (CNRS) CS80054, 8LinkoCare Life Sciences AB, 9College of Nursing and Children's Hospital Research Institute of Manitoba, Rady Faculty of Health Sciences, University of Manitoba, 10Health Policy Research Center, Institute of Health, Shiraz University of Medical Sciences

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Here we describe a protocol to investigate the prenylation and guanosine-5'-triphosphate (GTP)-loading of Rho GTPase. This protocol consists of two detailed methods, namely membrane fractionation and a GTPase-linked immunosorbent assay. The protocol can be used for measuring the prenylation and GTP loading of different other small GTPases.

Tags

Small GTPase Prenylation GTP Binding Membrane Fractionation GTPase-linked Immunosorbent Assay Rho GTPase Cellular Function Post-translational Modification Glioblastoma Cell Lines Simvastatin Sub-cellular Fractionation RhoA Confluency Cold PBS EDTA
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