Please help me fill this chart as well as show calculations/formulas to learn. Q

ID: 316322 • Letter: P

Question

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?

Explanation / Answer

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