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Detecting DNA Binding Protein Activity Using a Membrane-Based Approach

Natalie Betz
Promega Corporation
Publication Date: 2006


The SAM® Biotin Capture Membrane can be used to study biotinylated molecules based on their affinity for streptavidin. The SAM® Membranes bind linearly and with low non-specific interactions to substrates. For DNA:protein binding studies, they can be used with either biotin-labeled proteins and [32P]-labeled oligos or with biotinylated oligos and [35S]-labeled proteins. This method provides an easy and convenient nongel-based method for detecting the ability of an in vitro-expressed target protein to specifically bind to a DNA oligonucleotide and serves as an alternative to the classic gel mobility shift, electrophoretic mobility shift (EMSA), or gel retardation assays.

Promega Notes 93, 5–7.

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