Journal
JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY
Volume 23, Issue 6, Pages 931-945Publisher
INFORMA HEALTHCARE
DOI: 10.1080/14756360701810082
Keywords
Assay; miniaturization; combinatorial chemistry; cytotoxicity; anthracyclines; screening; HTS; fluorescence; resazurin; cell viability; NCEB1; cancer
Funding
- Commonwealth Foundation for Cancer Research
- NATIONAL CANCER INSTITUTE [P30CA008748] Funding Source: NIH RePORTER
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In response to the need for inexpensive high throughput assays for anti-cancer drug screening, a 1536-well microtiter plate based assay utilizing the Alamar Blue fluorescent dye as a measure of cellular growth was validated in 10L assay volume. Its robustness was assessed in a screen against a library of 2000 known bioactives; with an overall Z' value of 0.89 for assay robustness, several known cytotoxic agents were identified including and not limited to anthracyclines, cardiac glycosides, gamboges, and quinones. To further test the sensitivity of the assay, IC50 determinations were performed in both 384-well and 1536-well formats and the obtained results show a very good correlation between the two density formats. These findings demonstrate that this newly developed assay is simple to set up, robust, highly sensitive and inexpensive. It could potentially provide a rapid way to screen established and primary tumor cell lines against large chemical libraries.
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