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Effect of various reducing agents on the induction of protein splicing. Mixtures of MUNΔ and UCΔH, reconstituted in the absence of DTT, were supplemented with 1.25 mM glutathione (lane 2), 1.25 mM TCEP (lane 3), 1.25 mM DTT (lane 4), or no reducing agent (lane 5) and incubated at 25°C and pH 7.5 for 6 hr, followed by SDS/PAGE without additional reducing agent in the sample buffer and Western blot analysis with anti-His-tag antibody.
