Fluorescent Western blotting allows multiplex detection without stripping and reprobing for analytes, and can be used for quantitative detection. Various fluorescent conjugates can be employed depending on the instrument setup. For optimal detection, Alexa Fluor® 680 and Alexa Fluor® 790 conjugates can be used to achieve high sensitivity Western blots.
Near-Infrared (NIR) Fluorescent Conjugates
Antibodies conjugated with far-red- and infrared-emitting dyes are suitable for a variety of techniques requiring the highest sensitivity. Alexa Fluor® 680 and 790 dyes are more sensitive than visible light-emitting dyes due to lower fluorescence quenching of the conjugates and higher extinction coefficients of the dyes. Low sample autofluorescence in this region of the spectrum results in lower background signal compared with other fluorophores.
Alexa Fluor® 680 and 790 conjugates can be used for high sensitivity Western blots, quantitative Western blots, in-gel Western blots, microWestern arrays, in-cell Western assays, on-cell Western assays, and other techniques that require the brightest dyes.