Help me answer the questions
1. Provide a labeled figure showing the SDS-PAGE gel of the various fractions collected during
MGH expression and purification. Indicate the migrated placement of the molecular weight
standards with their appropriate molecular weight as tick marks in the margin of the gel
image. Generate a standard curve from the standards and estimate the molecular weight of
your purified MGH and other major polypeptides detected, denoting them with tick marks in
the gel image margin.
2. Show the standard curve and linear regression analysis for the protein assay done for
purified MGH. Provide an estimate on the final MGH concentration you purified.
3. Provide a labeled figure showing the immunoblot using anti-H6-HRP antibody. Generate a
standard curve from the standards and estimate the molecular weight of your immunodetected MGH.
4. Comment about the efficacy of the Ni2+-NTA resin to purify MGH from your E coli lysate.
What was the total yield of MGH by this procedure? How pure is your MGH preparation?
Compare the results of the immunoblot with the corresponding SDS-PAGE; what similarities
and significant differences did you see? Was there much detectable yellow-green color in
the unbound fraction?
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