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Direct the stream of water down the side of the vial rather than directly onto the peptide cake, and swirl gently rather than shaking
On ice, cells were scraped and collected in extraction buffer containing methanol:acetonitrile:H 2 O (4:4:2), 15 M glutaric acid and 0.5% formic acid
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Focusing on endogenous peptides, analysis of HEK 293T cell lysate tryptic digests by SCX chromatography coupled with pS-MIP enrichment resulted in the identification of 924 phosphopeptides in only 10 g of SCX fractionated peptide sample (starting material 40 g trypsinized cell lysate)
Setting realistic goals: Define specific objectives (pain reduction, return to specific activities, healing verified on imaging) rather than vague "feel better." Understand realistic timeline (weeks for soft tissue, months for bone, months to years for cartilage) preventing premature disappointment
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