IRDye Streptavidins for Detection of Biotin-labeled Molecules.
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IRDye® Streptavidins

Streptavidin binds to biotin very strongly and is used to detect biotinylated proteins. IRDye infrared dye-labeled Streptavidins can be used as a secondary detection reagent for microscopy, protein arrays, In-Gel Westerns, ELISA, and Western blotting. IRDye Streptavidins can also be used to detect Strep-tag. Imaging can be performed on Odyssey® Infrared Imaging Systems.

Streptavidin

Figure 1. Example of IRDye Streptavidin used for a protein array. Spots are Biotinylated BSA + 2-fold serial dilution of IRDye 680 Streptavidin. Data were obtained from a 21-micron scan on an Odyssey® Infrared Imaging System.

All IRDye streptavidins are supplied as a liquid in buffer containing 10 mM phosphate, 183 mM NaCl, 2.7 nM KCl, pH 7.4 with sodium azide 0.005% (w/v) as a preservative. To use, centrifuge briefly before use to eliminate aggregates that may have formed in solution. This will reduce non-specific background staining. A final concentration of 0.2 to 1.0 µg/ml (1:1,000 to 1:5,000) is usually satisfactory for most applications; however, appropriate dilution may need to be determined empirically. For membrane-based applications and In-Gel Westerns, it is recommended to add SDS (0.02% to 0.1% final concentration), in addition to Tween® 20 (0.1 to 0.2% final concentration) during the detection incubation step to reduce non-specific background staining.



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