Detecting DNA Binding Protein Activity Using a Membrane-Based Approach

The SAM^2®® Biotin Capture Membrane can be used to study biotinylated molecules based on their affinity for streptavidin. The SAM^2®® 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.