Related Experiment Video
Updated: Aug 18, 2026
Measuring G-protein-coupled Receptor Signaling via Radio-labeled GTP Binding
Published on: June 9, 2017
Quantification of absolute Ras-GDP/GTP levels by HPLC separation of Ras-bound [(32)P]-labelled nucleotides
Ignacio Rubio1, Rico Pusch, Reinhard Wetzker
1Research Unit Molecular Cell Biology, Medical Faculty, Friedrich-Schiller-University, Drackendorfer Str. 1, 07747 Jena, Germany. b5igru@rz.uni-jena.de
Abstract:
Ras guanine nucleotide binding protein (GTPase) activation is a widely assessed readout in cell biological studies. We describe an improved approach for the quantitative analysis of total GDP and GTP bound to Ras. The present method involves HPLC separation and online detection/quantitation of Ras-bound [(32)P]-labelled GDP/GTP. As compared to standard approaches that are time consuming and/or provide only semi-quantitative data, this technique allows the rapid processing of large numbers of samples for the quantitative determination of Ras-bound GDP and GTP.
More Related Videos
13:51Detection of Small GTPase Prenylation and GTP Binding Using Membrane Fractionation and GTPase-linked Immunosorbent Assay
Published on: November 11, 2018
09:26Identifying the Binding Proteins of Small Ligands with the Differential Radial Capillary Action of Ligand Assay (DRaCALA)
Published on: March 19, 2021
