1. You want to purify a protein using anion exchange column chromatography. In this technique, the solid state beads are positively charged. What charge would you want your protein to have in order for it to "stick" to the beads? Neutral Positive Negative 2. In your anion exchange experiment, your protein of interest has a pI of 6.0. Which of the following buffer solutions would you want to use when loading your protein extract onto the column so that your protein "sticks" to the beads? pH 4.5 pH 7.3 pH 6.0 3. You want to analyze your chromatography results using gel electrophoresis. If your protein is a pentamer consisting of 2 alpha subunits, 1 beta subunit, and 2 gamma subunits, how many bands would you expect to see on your gel? The alpha subunits are 32kDa. The beta subunit is 32kDa. The gamma subunits are 15kDa. 1 5 2 3 4. You want to analyze your chromatography results using gel electrophoresis. If your protein is a pentamer consisting of 2 alpha subunits, 1 beta subunit, and 2 gamma subunits. The alpha subunits are 32kDa. The beta subunit is 32kDa. The gamma subunits are 15kDa. Which subunit would you expect to find travelled the farthest on your gel (near the bottom of the gel)? Alpha & Beta Beta Alpha Gamma All five subunits travel together on the gel
1. You want to purify a protein using anion exchange column chromatography. In this technique, the solid state beads are positively charged. What charge would you want your protein to have in order for it to "stick" to the beads?
Neutral
Positive
Negative
2. In your anion exchange experiment, your protein of interest has a pI of 6.0. Which of the following buffer solutions would you want to use when loading your protein extract onto the column so that your protein "sticks" to the beads?
pH 4.5
pH 7.3
pH 6.0
3. You want to analyze your chromatography results using gel electrophoresis. If your protein is a pentamer consisting of 2 alpha subunits, 1 beta subunit, and 2 gamma subunits, how many bands would you expect to see on your gel? The alpha subunits are 32kDa. The beta subunit is 32kDa. The gamma subunits are 15kDa.
1
5
2
3
4. You want to analyze your chromatography results using gel electrophoresis. If your protein is a pentamer consisting of 2 alpha subunits, 1 beta subunit, and 2 gamma subunits. The alpha subunits are 32kDa. The beta subunit is 32kDa. The gamma subunits are 15kDa. Which subunit would you expect to find travelled the farthest on your gel (near the bottom of the gel)?
Alpha & Beta
Beta
Alpha
Gamma
All five subunits travel together on the gel
In anion exchange form of affinity column chromatography, since the solid state beads are positively charged, a negatively charged protein will interact with the matrix. In denaturing SDS-PAGE gel electrophoresis, the protein molecules travel according to their molecular weights. The mobility of the protein in SDS-PAGE is inversely proportional to the molecular weights.
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