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Detecting DNA-Binding Protein Activity using a Membrane-Based
Approach
Natalie Betz, Ph.D.
Promega Corporation
The SAM2® Biotin Capture Membrane can be used to study biotinylated
molecules based on their affinity for streptavidin. The SAM2® 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.
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