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

Javad Alizadeh; Shahla Shojaei; Simone da Silva Rosa; Adel Rezaei Moghadam; Amir A. Zeki; Mohammad Hashemi; Marek J. Los; Joseph W. Gordon; Saeid Ghavami

doi:10.3791/57646

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Rilevazione di piccolo GTPase prenilazione e GTP Binding mediante frazionamento di membrana e GTPase-collegata dell'immunosorbente

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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

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Rilevazione di piccolo GTPase prenilazione e GTP Binding mediante frazionamento di membrana e GTPase-collegata dell'immunosorbente

9,650 Views

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13:51 min

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November 11, 2018

DOI:

10.3791/57646-v

DOI:

10.3791/57646-v

•

13:51 min

November 11, 2018

•

8 Views

Javad Alizadeh^1,2, Shahla Shojaei^1,2,3, Simone da Silva Rosa¹, Adel Rezaei Moghadam^1,2, Amir A. Zeki⁴, Mohammad Hashemi⁵, Marek J. Los^6,7,8, Joseph W. Gordon^1,9, Saeid Ghavami^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

Summary

Qui descriviamo un protocollo per studiare la prenilazione e guanosina-5'-trifosfato (GTP)-caricamento di Rho GTPasi. Questo protocollo è costituito da due metodi dettagliati, vale a dire del kit membrana frazionamento e un GTPase-collegata dell'immunosorbente. Il protocollo può essere utilizzato per misurare la prenilazione e GTP caricamento di diverse altre piccole GTPasi.

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Rilevazione di piccolo GTPase prenilazione e GTP Binding mediante frazionamento di membrana e GTPase-collegata dell'immunosorbente

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Summary

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