Experimental Molecular Weight higher than Literature Value

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SUMMARY

The calculated molecular weight of alkaline phosphatase after purification is 100,000 g/mol, significantly higher than the literature value of 89,000 g/mol. The discrepancy may be attributed to the presence of EDTA, which can inhibit the enzyme and potentially increase the measured molecular weight. The discussion highlights the importance of purification methods, such as gel electrophoresis and dialysis, in obtaining accurate molecular weight measurements. Further details on the purification process are necessary to provide a definitive explanation for the observed results.

PREREQUISITES
  • Understanding of alkaline phosphatase and its properties.
  • Knowledge of gel electrophoresis techniques.
  • Familiarity with dialysis methods for protein purification.
  • Basic principles of molecular weight determination.
NEXT STEPS
  • Research the effects of EDTA on alkaline phosphatase activity and molecular weight.
  • Learn about gel electrophoresis protocols for protein purification.
  • Investigate dialysis techniques and their impact on protein purity.
  • Explore methods for accurately determining molecular weight of proteins.
USEFUL FOR

This discussion is beneficial for biochemists, molecular biologists, and laboratory technicians involved in protein purification and analysis, particularly those working with alkaline phosphatase.

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Homework Statement


I just purified alkaline phosphatase, and my calculated molecular weight is 100000 g/mol, while literature value is 89000 g/mol. According to how others in my lab did calculations, my calculations are performed correctly, so I am left to explain why my purification is higher than expected.

Homework Equations


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The Attempt at a Solution


I was thinking that since alkaline phosphatase can be inhibited by EDTA, my "purified" enzyme contains some EDTA, making the molecular weight higher.
 
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Did you purify it by gel electrophoresis? What was the supporting electrolyte? Did you completely dialyze your product?
 
You do not give any indication of what method used for getting the MW so what can anyone say?
 

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