title: Collecting data to determine the Ca²⁺-binding properties of DM-nitrophen and proteins.
abstract: Conventional techniques used to measure Ca(2+) binding are too slow to determine accurately the fast binding kinetics of most molecules such as Ca(2+)-binding proteins (CBPs). We have developed an ultra-fast in vitro technique for measuring the Ca(2+)-binding properties of CBPs following flash photolysis of caged Ca(2+). Although the details of the setup, the mathematics, and the analysis involved in this technique have been published elsewhere, many of the practical details regarding the actual measurements have, until now, only been described minimally. Here, we present a protocol to gather the data necessary to determine the kinetic properties of a caged-Ca(2+) compound and a CBP.
authors.affiliation: Department of Neurology, David Geffen School of Medicine, UCLA, Los Angeles, California 90095;
authors.forename: Guido C
authors.initials: GC
authors.lastname: Faas
authors.affiliation: Department of Neurology, David Geffen School of Medicine, UCLA, Los Angeles, California 90095; Department of Physiology, David Geffen School of Medicine, UCLA, Los Angeles, California 90095.
authors.forename: Istvan
authors.initials: I
authors.lastname: Mody
mesh_terms: D000085:Acetates / Q000737:chemistry / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D005029:Ethylenediamines / Q000737:chemistry / Q000378:metabolism*; D011506:Proteins / Q000737:chemistry / Q000378:metabolism
chemical_list: D000085:Acetates; D002135:Calcium-Binding Proteins; D005029:Ethylenediamines; D011506:Proteins; C057014:DM-nitrophen