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J Biomol Screen. 2016 Dec;21(10):1125-1131. doi: 10.1177/1087057116652895. Epub 2016 Jul 10.

A Platform to Enable the Pharmacological Profiling of Small Molecules in Gel-Based Electrophoretic Mobility Shift Assays.

Author information

1
1 Division of Preclinical Innovation, National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD, USA.
2
Primary Pharmacology Group, Pharmacokinetics, Dynamics & Metabolism-New Chemical Entities, Pfizer, Groton, CT, USA.
3
Assay Development and Drug Repurposing Core Facility, Department of Pharmacology & Toxicology, Michigan State University, East Lansing, MI, USA.
4
2 Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, CA, USA.
5
3 Luxon Engineering, LLC, San Diego, CA, USA.

Abstract

We describe a polyacrylamide gel casting cassette that overcomes limitations of commercially available gel electrophoresis equipment. This apparatus molds a single polyacrylamide gel that can evaluate more than 200 samples in parallel, is loaded with a multichannel pipettor, and is flexible with respect to composition of the separating matrix. We demonstrate its use to characterize inhibitors of enzymes that modify protein and nucleic acid substrates. Throughputs of greater than 1000 samples per day were achieved when this system was paired with a quantitative laser-based imaging system, yielding data of remarkable quality.

KEYWORDS:

electrophoretic mobility shift assay (EMSA); electrophoretic techniques; enzymatic assays and analyses; high-throughput screening (HTS); polyacrylamide gel electrophoresis (PAGE); small-molecule testing

PMID:
27269812
PMCID:
PMC5269537
DOI:
10.1177/1087057116652895
[Indexed for MEDLINE]
Free PMC Article

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