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Centre for Cell Signalling and Molecular Genetics 8M Urea Lysis buffer • 1M dithiothreitol(DTT)1ml aliquot DTT: 0.1542 g ddH20 1 ml • Notes: – For Weigh urea into 50ml falcon, then add 30mls ddH20, and then the glycerol, 1M DTT and 1.5M Tris. – Allow urea to dissolve through gentle agitation, before adding the 20% SDS, and finally topping up to 50mls with ddH20. – Do not add PMSF (1/100) until just before lysis. – **For BSA/Bioradprotein assays, DTT will interact with the assay. Therefore we suggest cutting DTT to 1/1000 for the assay itself to, albeit far from perfect, improve assay reliability.