b) Mix the cell lysate with SDS loading buffer to make the desired final concentration.Measure protein concentration using cell lysis-compatible protein assay (BCA protein assay). Dilute the supernatant lysate and the whole lysate without centrifugation at 1:4, 1:8 and 1:16 with 1XPBS. Then collect supernatant into an appropriately labeled tube (for the detection of membrane bound proteins, use the insoluble, cell pellet).
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