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Comparison of EMSA Detection Methods

Comparison of Infrared Fluorescent EMSA and other methods:

IRDye® Infrared Fluorescence Dyes Radioisotopes Biotin/Streptavidin

(Such as Thermo Scientific LightShift)

Total Time: 1.5h Total Time: 4.5 – 24h Total Time: 4.5 – 5h
Easy access and disposal Regulatory restrictions, disposal hassles, and cost
Fluorescent oligos have extended stability Short half-life of labeled oligos Chemiluminescent signal is unstable
Non-hazardous Hazardous
Wet gel imaged without removing gel plates Gel drying and film/phosphor-screen exposure required Membrane transfer is required
Fast, convenient, direct detection Time-consuming, inconvenient detection Indirect detection method. Blocking, streptavidin incubation, and washes are required
Gel can be replaced and run longer, if needed Gel run time cannot be extended Gel run time cannot be extended
Results available in less than 2 hours Results typically not obtained until the next day Detection steps add several hours to protocol
time_comparison

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