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In this case, 0.5 mg

Discarding the upper plasma, wash with three times volume of PBS for 3 times (re-suspend blood cell with PBS, centrifuge at 600 g for 10 minutes), add equal volume of Reagent I

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In experiments aiming to capture more interacting proteins, the beads were washed five times with 0.5 ml of cold lysis buffer (50 mM Tris-HCl, 0.8% Igepal-CA630, 5% glycerol, 150 mM NaCl, 1.5 mM MgCl2, pH 7.5, 1 mM DTT, cOmplete mini protease EDTA-free inhibitor) containing additional 0.2% NP-40, referred to as soft wash in this manuscript

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