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Formation of mixed tetramers. A 9−12% gradient, non-denaturing polyacrylamide gel of the mixed tetramer is used to visualize the fluorescent-labeled proteins under UV excitation. Lanes 2−6 result from anion exchange chromatography as described in the text. Lane 1: wild type BsPFK-Texas Red, lane 2: fraction 5, lane 3: fraction 19, lane 4: fraction 25, lane 5: fraction 33, lane 6: fraction 42, lane 7: K90,91EAlexaFluor488, lane 8: dimer exchange mixture following dialysis.
