Open AccessA 1536 Colorimetric SPAP Reporter Assay: Comparison with 96- and 384-Well Formats
Author(s) -
J. C. W. Comley,
Tony Reeves,
Philip Robinson
Publication year - 1998
Publication title -
slas discovery
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.907
H-Index - 75
eISSN - 2472-5560
pISSN - 2472-5552
DOI - 10.1177/108705719800300308
Subject(s) - reporter gene , microbiology and biotechnology , bioreporter , colorimetry , bioassay , chemistry , biology , gene , gene expression , chromatography , biochemistry , genetics
We report the successful miniaturization of a functional cell-based reporter gene assay. Utilizing interleukin-1beta (IL-1/β)-induced secreted placental alkaline phosphatase (SPAP)-catalyzed colorimetric readout, we reduced the assay volume to 10 μl using a Greiner 1536-well microplate. Our experiences of assay development, liquid handling (using a Hydra® 96; Robbins Scientific, Sunnyvale, CA), and detection (using the SpectraImage and SpectraFluor-Plus plate readers, Tecan Austria GmbH, Grodig, Austria) in 1536 wells are discussed. The effect of a set of 1,280 compounds in this SPAP reporter assay were compared between 96-, 384-, and 1536-well formats and were shown to be very similar. We conclude that cell-based reporter gene assays using SPAP-catalyzed color readouts are sensitive and highly reproducible in 1536-well plates and should be considered as a cost-effective alternative to luciferase reporters for miniaturized assay formats. Finally, we review the prospects for the implementation of routine HTS in 1536-well plates based around the instrumentation investigated.