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J Pharmacol Exp Ther (2004) 308(2):5218

see appendix A)

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In brief, 50 g protein aliquots from each sample were mixed with denaturing buffer (8 M Urea, 0.1 M TrisHCl pH 8.5), then each aliquot was reduced and alkylated via sequential 20-min incubations with 5 mM TCEP (tris(2-carboxyethyl)phosphine) and 10 mM iodoacetamide at room temperature in the dark while being mixed at 1200 rpm in an Eppendorf thermomixer