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The reagents in different kits cannot be mixed with each other

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That means the practical answer to how long is bac water good for after opening is usually up to 28 days, provided the original expiration date has not arrived earlier and the vial still meets visual and handling checks

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2) Resuspend the cells in 10 mL of Equilibration buffer (LE buffer) with appropriate amount of PMSF (Final working concentration can be 1mM) and add lysozyme (Final working concentration is 0.2-0.4 mg/mL), or other protease inhibitors cocktail added