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Binding of LC/A to the recombinant SNAP-25. SNAP-25 was coated in an ELISA plate and incubated with the indicated 3xFLAG-LC/A derivative for 1 h at room temperature. Bound LC/A was detected with mouse α-FLAG-HRP antibody (Pierce, 1:10,000 final dilution) for 1 h at room temperature and developed with Ultra TMB (Pierce) for 20 min at room temperature. The reaction was quenched with H2SO4, and A450 was determined. The amount of LC/A needed to bind 50% SNAP-25 was determined.
