Homework SoursePlease help me fill this chart as well as show calculationsf
Please help me fill this chart as well as show calculations/formulas to learn. Question is from the filled chart, which fraction had most enzyme?
First column:
Absorbance 420nm
Protein mix 2.332
Fraction 1 0.236
Fraction 2 0.127
Fraction 3 0.070
Second column:
Absorbance 280nm
Protein mix 0.370
Fraction 1 0.115
Fraction 2 0.150
Abs Abs Total Specific activity yield Fold 280 protein (mg) For alkaline phosphatase purification (umol/min/mg) nm nm Protein A mix Fraction 1 Fraction 2 Fraction 3 Assume the extension coefficient of all these protein at 280 nm is 1 mg/mL/cm. Assume alkaline phosphatase\'s product has an extension coefficient of 0.1 umol/mL/cm Which fraction has the most of enzyme? Solution
Total protein (mg/ml) = Absorbance at 420nm and 280nm /standard
Alkaline phosphatase activity = Absorbance at 420nm/standard*10
Abs
420nm
Abs
280nm
total protein
(mg/ml)
Specific activity for Alkaline phosphatase
% yield
0.236
0.115
0.325602
0.932548
93.2548
0.127
0.15
0.134334
0.544597
54.45969
0.07
0.182
0.061024
0.300172
30.01715
More enzyme activity (93.25%) is seen in the Absorbance at 420nm.
| Abs 420nm | Abs 280nm | total protein (mg/ml) | Specific activity for Alkaline phosphatase | % yield |
| 0.236 | 0.115 | 0.325602 | 0.932548 | 93.2548 |
| 0.127 | 0.15 | 0.134334 | 0.544597 | 54.45969 |
| 0.07 | 0.182 | 0.061024 | 0.300172 | 30.01715 |
