binding proteins by
-ligand overlay. Proteins were dot-blotted onto nitrocellulose and then incubated with 45CaCl2 in a calcium overlay assay . 45Ca-labeled proteins were visualized by autoradiography (7-d exposure). 1, equine cytochrome c, used as negative control (5 μg); 2, purified M. loti 20 kDa Stains-all blue-staining protein (5 μg); 3, rabbit calsequestrin, used as positive control (1 μg).