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Briefly describing, 450 l phosphate buffer was added to 50 l supernatant and was subject to ultrafiltration (ULTRAFREE-0.5 Centrifugal Filter Device (UFV5BGC100), Millipore, MA, USA) at 12000 g at 4C for 60 min
Repeated freeze-thaw cycles degrade the peptide
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Also, limit alcohol and be cautious with over-the-counter appetite suppressants
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