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NAADP and other Ca(2+)-mobilizing messengers are membrane impermeant and thus must be added directly to cell-free or broken-cell preparations to effect Ca(2+) release. The sea urchin egg homogenate, where the biological activity of NAADP was first reported, remains the gold standard cell-free system for studying NAADP-mediated Ca(2+) release. Here we describe how to prepare sea urchin egg homogenate and use it to measure NAADP-mediated Ca(2+) release.

Original publication

DOI

10.1101/pdb.prot076901

Type

Journal article

Journal

Cold Spring Harb Protoc

Publication Date

02/09/2014

Volume

2014

Pages

988 - 992

Keywords

Adenosine Triphosphate, Aniline Compounds, Animals, Calcium, Calcium Signaling, Cell Fractionation, Female, Inositol 1,4,5-Trisphosphate, Male, NADP, Ovum, Sea Urchins, Xanthenes