2009-09-11_-_Lecture_4

2009-09-11_-_Lecture_4 - Biochemistry 461 FS 2009 Michigan...

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Biochemistry 461 FS 2009 Michigan State University Proteins: Primary Structure and Evolution September 11, 2009 Lecture 4 Quantitatively following a protein purification: • At each step (crude extract or following a particular procedure), measure the volume of solution and assay: Enzyme activity in units/mL; Protein concentration in mg/mL (Bradford assay or similar). Total protein = (protein concentration)*(volume) Total activity = (units/mL)*(volume) Specific activity = (total activity)/(total protein) • For the crude extract, these three measurements are considered the “initial” values. For subsequent steps: Yield = (total activity)/(initial total activity) * 100 % Fold purification = (specific activity)/(initial specific activity). Protein Purification: Following Progress 2
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Protein Purification: Following Progress • Activity is a measure of the amount of enzyme . • Specific activity is a measure of the
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2009-09-11_-_Lecture_4 - Biochemistry 461 FS 2009 Michigan...

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