High Throughput Gel Shifts

To determine whether a given transcription factor activity is present in a sample, you need look no farther than the standard electrophoretic mobility shift assay (EMSA). In an EMSA, researchers mix a radioactively labeled DNA probe with a protein extract and run the entire reaction on a nondenaturing polyacrylamide gel. Because the protein-bound probe will migrate more slowly than a free probe, the experiment is described as a "gel shift." Unfortunately, the gel shift is a cumbersome way to pro

Written byJeffrey Perkel

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Using the array is simple. Researchers mix protein extracts (e.g., nuclear extracts [25 µg]), with prelabeled probe mix supplied by Panomics. This mix contains all of the cis elements the MYRIA array can currently detect, each of which was selected because of its wide representation in published literature. After an incubation period, the investigator resolves the free and bound complexes on an agarose gel and purifies the bound complex. Then, the user interrogates the MYRIA array with this purified probe, on which each element is spotted in duplicate and at two separate concentrations.

Currently, the system is supplied with biotin-labeled probes and nylon membrane-based arrays. Panomics is in the process of developing a glass slide-based system, using fluorescently labeled probes to allow for dual-color analyses. This feature would allow researchers to compare the effect of drug treatment on a single cell line, for example. According to Jason Li, Panomics president ...

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