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Preparation of whole cell extracts for Nrf2 and Keap 1 western blot Neutrophils were pelleted, washed twice with ice-cold PBS and resuspended in 100l of ice-cold lysis buffer [50 mM Tris-HCl (pH 6.8), 10% (vol/vol) glycerol, and 2% (wt/vol) SDS]