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This observation, together with the fact that 1 equivalent of DTT is not consumed in the reaction, indicated that intramolecular amide formation by nucleophilic attack of the AX side chain amino group to its electrophilic -lactam carbonyl, under these conditions, occurs only in the presence of DTT

concentrations drop by more than half between age 20 and 60

Generally, its best to use gentler cooking methods

Therefore, AX reactivity and conjugation to carrier molecules have been the subject of thorough investigation ( Initial assays preincubating proteins with reducing agents before treatment with AX showed a clear decrease in AX-protein adduct formation in the presence of DTT, whereas the effect of GSH was less consistent ( Figure 1 )

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