TY - JOUR
AU - Ward, Thomas R.
AB - The biotin‐binding protein streptavidin exhibits a high stability against thermal denaturation, especially when complexed to biotin. Herein we show that, in sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE), streptavidin is stabilized at high temperature in the presence of biotinylated fluorescent probes, such as biotin‐4‐fluorescein, which is incorporated within the binding pocket. In nondenaturing SDS‐PAGE, streptavidin is detectable when complexed with biotin‐4‐fluorescein using a UV‐transilluminator. Using biotin‐4‐fluorescein, the detection limit of streptavidin lies in the same range as with Coomassie blue staining. The functionality of streptavidin mutants can readily be assessed from crude bacterial extracts using biotin‐4‐fluorescein as a probe in nondenaturing SDS‐PAGE.
TI - Electrophoretic behavior of streptavidin complexed to a biotinylated probe: A functional screening assay for biotin‐binding proteins
JF - Electrophoresis
DO - 10.1002/elps.200406148
DA - 2005-01-01
UR - https://www.deepdyve.com/lp/wiley/electrophoretic-behavior-of-streptavidin-complexed-to-a-biotinylated-GO3zVEEDqb
SP - 47
EP - 52
VL - 26
IS - 1
DP - DeepDyve
ER -