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Repeated freeze-thaw cycles and repeated stopper punctures both shift the reduced to oxidised balance, so aliquoting matters more for glutathione than for a non-thiol compound

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Proteins were then transferred onto a polyvinylidene difluoride (PVDF) membrane using an iBlot2 PVDF stack (Invitrogen) and blocked with 5% bovine serum albumin in tris-buffered saline with 0.1% Tween-20 (TBST) for 1 hour at room temperature