Ligand binding with stopped-flow rapid mixing.
Stopped-flow rapid mixing is a common, direct technique for the study of ligand-binding reactions. In this method, protein and ligand are mixed together at relatively high velocities directly into an observation chamber, so that time courses for reactions occurring on time scales as short as a few milliseconds can be measured. This chapter presents an introduction to this technique, including a discussion of experimental and technical issues that must be addressed when designing stopped-flow experiments. Simple experiments for measuring reaction dead time and flushing volume are also described along with the details of several common reaction schemes and methods for data analysis.
Hargrove
Mark S
MS
Department of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, IA, USA.
D000818:Animals; D002248:Carbon Monoxide / Q000378:metabolism; D006736:Horses; D066298:In Vitro Techniques; D007700:Kinetics; D008024:Ligands; D008967:Molecular Biology / Q000295:instrumentation / Q000379:methods*; D009211:Myoglobin / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism*; D019810:Sodium Azide / Q000378:metabolism
D008024:Ligands; D009211:Myoglobin; D011506:Proteins; D002248:Carbon Monoxide; D019810:Sodium Azide