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Journal of Biomolecular Screening
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1087057107301522v1
12/5/705    most recent
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*Gene*GEO Profiles
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*CALCIUM COMPOUNDS
*CALCIUM, ELEMENTAL
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Article

Evaluation of No-Wash Calcium Assay Kits: Enabling Tools for Calcium Mobilization

Hong Xin*, Yuanping Wang, Matthew J. Todd, Jianshen Qi, Lisa K. Minor

Johnson & Johnson PRD, U.S. RED

* To whom correspondence should be addressed. E-mail: hxin{at}prdus.jnj.com.


  Abstract
The no-wash calcium assay kits developed by Molecular Devices Corporation have greatly enhanced throughput of cell-based calcium mobilization HTS assays, and enabled screening using non-adherent cells. The FLIPR Calcium 3 Assay Kit, optimal for targets that have proteins or peptides as agonists, has two potential drawbacks: 1) a significant downward spike in fluorescence signal upon liquid transfer that can be the same magnitude as the agonist response making data analysis difficult; 2) medium removal is required for some targets which essentially re-introduces a wash step. Several no-wash products were introduced in 2005. We have compared the Fluo-4 NW Calcium Assay Kit, and the BDTM Calcium Assay Kit with the FLIPR Calcium 3 Assay Kit using human native rhabdomyosarcoma cells expressing the urotensin-II (U-II) receptor (UT). The BDTM Calcium Assay Kit gave the best performance in the true no-wash mode, where both agonist and antagonist activity could be easily quantified. Although these new products provide us with additional options for measuring calcium mobilization, the different results observed with each kit, using UT receptor as an example, suggest that one should characterize all dyes against each target in a systematic way prior to choosing one for HTS.

First published on May 30, 2007, doi:10.1177/1087057107301522

Journal of Biomolecular Screening 2007;12:705.

A more recent version of this article appeared on August 1, 2007

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[Abstract] [PDF]