User:Douglas M. Fox/Notebook/AU CHEM-571 F2011 Lab Support/2014/10/14

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SDS-PAGE #2 Mix 10 μL 0.6 g/L lysozyme with 10μL SDS-PAGE sample buffer in 1.5 mL centrifuge vial Mix 10 μL 0.12 g/L lysozyme with 10μL SDS-PAGE sample buffer in 1.5 mL centrifuge vial Mix 10 μL 30:1 Au/lysozyme colloid with 10μL SDS-PAGE sample buffer in 1.5 mL centrifuge vial Mix 10 μL 0.12 g/L unknown protein with 10 μL SDS-PAGE sample buffer in 1.5 mL centrifuge vial Place in heating block (set at 90 °C) for 5 minutes Store in refrigerator overnight One group prepare additional 1 L of running buffer from 10X concentrate Note: Ideal concentration for pure protein is 0.5 - 4.0 μg (40 - 60 μg for crude samples). Using 10 μL of 0.12 g/L will load 1.2 μg protein into the well if entire 20 μL is used. Protein Dialysis Continue dialysis from previous tasklist Buffer Preparation Prepare 30 - 40 mL 66 mM potassium phosphate buffer, pH 6.24 Use HPLC water 8.98 mg/mL KH2PO4 should produce pH 4 - 4.1 Add 1 M KOH to adjust pH to 6.24. To avoid this step a mixture of monobasic and dibasic potassium phosphate was used. Store