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Calcium-Binding Protein Protocols Calcium-Binding Protein Protocols

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Spatial Distribution of Ca 2+ -<strong>Binding</strong> <strong>Protein</strong>s 387<br />

Fig. 1. Purification of singly labeled FL-calmodulin. The FL-calmodulin reaction<br />

mixture after PD-10 filtration was analyzed by HPLC on a semipreparative Vydac<br />

C18 reverse phase column. The mixture shows three peaks in the following order:<br />

(a) unlabeled calmodulin; (b) singly labeled FL-calmodulin; (c) doubly labeled<br />

FL-calmodulin. The illustrated section of the chromatograms represent the 40–60%<br />

acetonitrile in the eluting solvent gradient. Absorbance (top panel) was measured at<br />

215 nm and fluorescence (bottom panel) was measured with transmission peaks of<br />

450 nm (excitation) and 528 nm (emission).

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