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Proteins were then precipitated by addition of 500 l methanol (pre-cooled at 80 C), 400 l LCMS-grade water, and 100 l chloroform, and the mixture was vortexed and centrifuged (10,000 g, 10 min, 4C) to afford a protein disc at the interface of CHCl3 and aqueous layers

Light and Oxygen Exposure:Minimize exposure to light and oxygen, as both can degrade peptides

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