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Author: Garrick Wilson Cell lysate preparation To 10mls of Brij + Ca2+/Mg2+ add 1 complete protease inhibitor tablet and dissolve at room temperature Then place on ice to cool down Wash cells 2x in PBS Add cold lysis buffer ~500ul/well of a 6 well plate Place plate on ice for 30min Harvest cells into eppendorf and centrifuge at 4˚C for 30mins at 15000rpm Remove supernatant into a clean eppendorf and store at -20˚C Pellet can also be retained and stored at -20˚C Quantify total protein lysates prior to use using the BCA assay kit