GFP was expressed in 60 ml E. coli culture and a 5 ml aliquot was applied to Strep-Tactin Superflow Plus. Protein was eluted using 2.5 mM desthiobiotin, giving a total protein yield of 3.1mg. C: Cleared lysate; E: Elution fractions; F: Flow-through; M: Markers; W: Wash.
Purified Strep-tagged thioredoxin (10 ng) was mixed with crude cell lysate from mammalian (NIH-3T3), insect (Sf9), and bacterial (E. coli) cells (each 10 µg total protein). After electrophoresis and western transfer, the Strep-tagged protein was detected using the Strep-tag Antibody and chemiluminescent detection.