Complete the Protein Purification table based on given data (show works). Volume
ID: 83020 • Letter: C
Question
Complete the Protein Purification table based on given data (show works).
Volume (mL) Step Activity [Protein] (mglml) 12000 205 Initial extract 12.85 216 5000 4.97 Ppt pH 166 After butanol 4450 0.18 12.42 12360 60% acetone ppt 11.88 26250 20 35-48% acetone ppt 5.70 18900 40-50% acetone ppt 20 0.67 7340 After charcoal 1.03 14200 10 40-48% acetone ppt Total Protein Total Specific Activity Yield Purification Fold Activity Purification Volume Activity (malmly Protein (umol min Step or (umol min (mL) mg) Recovery fraction (mg) (umol min) or (UnitsExplanation / Answer
mol/min or Units(U)
Total protein (mg) – the primary measurement is of protein concentration, i.e. mg/ml which is obtained using a protein assay. The total protein is calculated by multiplying the protein concentration with the total volume (i.e. mg/ml x ml = mg).
Total activity (units) – the activity, in units ml-1, is obtained from an activity assay. The total activity is calculated by multiplying the activity by the total volume (i.e. units/ml x ml = units).
Specific Activity – specific activity is a way to measure how much of a measured protein there is with all of the other contaminating proteins. Divide the Activity by the mg of protein.The higher this value, the higher purity.
Purification (fold) – refers to the number of multiples of a starting value. The purification “per step” can also be obtained by dividing the specific activity after that step by the specific activity of the material before that step.
• Yield (%) – The activity of the original homogenate is arbitrarily assumed to be . The yield (%) is calculated from the total activity (units) at each step divided by the total activity (units) in the homogenate, multiplied by 100.
A B C D E F G H I Purification Step or fraction Volume (ml) Total activity Activity (mol/min or Units(U)
) Protein (mg/ml) Total Protein (mg) Specific Activity(mol/min/mg) % yield % recovery Purification fold Initial extract 12000 205 24,60,000 12.85 1,50,000 1,542 100 (1) Ppt pH 5000 216 10,80,000 4.97 24,850 43.46 43.9 0.02 After butanol 4450 166 7,38,700 0.18 801 922.22 30 1.73 60% acetone ppt. 55 12360 6,79,800 12.42 683.1 995.17 27.6 1.60 35-48% acetone ppt. 20 26250 5,25,000 11.88 237.6 2209.59 21.3 0.72 40-50% acetone ppt. 20 18900 3,78,000 5.70 114 3315.79 15.3 0.48 After charcoal 32 7340 2,34,880 0.67 21.44 10955.22 9.54 0.41 40-48% acetone ppt. 10 14200 142000 1.03 10.3 13786.41 5.7 0.12Related Questions
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