Question

In a biology lab course students are asked to purify a 25 kD protein IUP using affinity chromatography . After going through the steps everyone performs gel electrophoresis using SDS-PAGE to check whether they have succeeded in purifying their proteins. The groups' gel is shown with different lanes corresponding to a molecular weight ladder, different fractions collected during the purification process, and crude cell lysate sample. Based on the SDS-PAGE data select whether the purification was successful or not, what mistake was made during the purification if any, and what the group's next step should be. Molecular weights (kDa) MW ladder Flow-through Wash fractions Elution fractions Crude cell lysate No mistakes, purification is successful Forgot Washes before elution, Add elution fractions to a fresh column and wash before eluting Did not add correct elution buffer,Elute protein from column using correct buffer Protein not present in lysate, Start over with lysate containing over-produced protein

          In a biology lab course students are asked to purify a 25 kD protein IUP using affinity chromatography . After going through the steps everyone performs gel electrophoresis using SDS-PAGE to check whether they have succeeded in purifying their proteins. The groups' gel is shown with different lanes corresponding to a molecular weight ladder, different fractions collected during the purification process, and crude cell lysate sample.

Based on the SDS-PAGE data select whether the purification was successful or not, what mistake was made during the purification if any, and what the group's next step should be.

Molecular weights (kDa)
MW ladder
Flow-through
Wash fractions
Elution fractions
Crude cell lysate

No mistakes, purification is successful
Forgot Washes before elution, Add elution fractions to a fresh column and wash before eluting
Did not add correct elution buffer,Elute protein from column using correct buffer
Protein not present in lysate, Start over with lysate containing over-produced protein

$In a biology lab course students are asked to purify a 25 kD protein IUP using affinity chromatography . After going through the steps everyone performs gel electrophoresis using SDS-PAGE to check whether they have succeeded in purifying their proteins. The groups' gel is shown with different lanes corresponding to a molecular weight ladder, different fractions collected during the purification process, and crude cell lysate sample. Based on the SDS-PAGE data select whether the purification was successful or not, what mistake was made during the purification if any, and what the group's next step should be. Molecular weights (kDa) MW ladder Flow-through Wash fractions Elution fractions Crude cell lysate No mistakes, purification is successful Forgot Washes before elution, Add elution fractions to a fresh column and wash before eluting Did not add correct elution buffer,Elute protein from column using correct buffer Protein not present in lysate, Start over with lysate containing over-produced protein$

Added by Carol A.

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