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The lysate was clarified by centrifugation at 15,000 nx g for 15 min, protein concentrations were determined using the BCA total protein assay (Pierce), and then appropriate volumes of the lysates were diluted with 2x reducing Laemmli buffer and heated for ten min at 95 C

This matters because it shows the legitimate pathway peptides can take when they go through proper clinical trials

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The sweet spot for most researchers using a 5mg vial is 1.0mL of bacteriostatic water

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