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Upptäckt av små GTPase Prenylation och GTP bindande använder membran fraktionering och GTPase-linked Immunosorbent Assay
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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
Chapters
- 00:00Title
- 01:39Determination of RhoA Localization Using Membrane/Cytosol Fractionation
- 06:05Measurement of RhoA GTP Load using Small G-protein Activation Assay
- 12:37Results
- 13:09Conclusion
Här beskriver vi ett protokoll för att undersöka prenylation och guanosin-5'-trifosfat (GTP)-lastning av Rho GTPase. Detta protokoll består av två detaljerade metoder, nämligen membran fraktionering och en GTPase-linked immunosorbent assay. Protokollet kan användas för att mäta prenylation och GTP lastning av olika andra små GTPases.
Tags
Small GTPaseRho GTPasePrenylationGTP BindingMembrane FractionationGTPase-linked Immunosorbent AssayGlioblastoma Cell LinesSimvastatinSubcellular FractionationRhoAPost-translational ModificationCell FateCellular ResponseCellular MotilityCell CycleProtein RegulationEDTAUltracentrifugationCytoplasmic FractionMembrane Fraction
