Determination of Molecular Weight of Proteins by SDS-Polyacrylamide Gel Electrophoresis
1.) If you chromatograph an oblong protein, it appears to be larger than a spherical protein of equal mass. Explain.
2.) Explain two advantages of using Kd values instead of calculating raw elution volumes of our protein on your gel.
3.) Explain how to determine exclusion limit of an unknown gel.
4.) What is the possible range for a Kd value? How does the value relate to the size of the protein?
5.) Why are gel filtration columns usually long and narrow while ion-exchange columns are short and fat?
6.) Can you use Sephadex G-75 to separate alcohol dehydrogenase (MW 150,000) from Beta-amylase (MW 200,000)? Explain your answer.
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