Topic 5: Protein Purification Techniques
Isolation and analysis methods, chromatography, electrophoresis.
Types and Descriptions of Column Chromatography
Click to show Types and Descriptions of Column Chromatography example problem
7da7
Match each of the following types of column chromatography with their corresponding descriptions.
Note: Each choice will be used exactly once.
| Your Choice | Prompt | |
|---|---|---|
| 1. ion exchange column (IEX) | ||
| 2. gel filtration column (GFC) | ||
| 3. reverse phase column (RPC) | ||
| 4. affinity column (AC) |
Drag one of the choices below:
- A - separate by charge, particle binding is controlled by pH of the solvent mobile phase
- B - separates molecules by differences in size as they pass through a porous particles in a column
- C - hydrophobic-based separations for protein characterization
- D - a specific ligand is attached to the particle resin in the column
Cell Disruption Techniques Identification
Click to show Cell Disruption Techniques Identification example problem
126a
Which one of the following cell disruption techniques correspond to the description 'Cells are pulverized through a combination of extreme cold.'.
Types of Column Chromatography
Click to show Types of Column Chromatography example problem
8482
Which one of the following types of column chromatography correspond to the description 'for one type, gradients of increasing organic solvent concentration are used to affect separations'.
Identification of Macromolecules in Gel Electrophoresis Processes
Click to show Identification of Macromolecules in Gel Electrophoresis Processes example problem
eca0
Which one of the following types of macromolecules correspond to the gel component or process 'sodium dodecyl sulfate (SDS)'.
Determining Net Charge of Proteins at Given pH
Click to show Determining Net Charge of Proteins at Given pH example problem
78e4
Isoelectric Point Problem
| Protein Name | isoelectric point (pI) | molecular weight |
|---|---|---|
| Succinate Ligase (SL) | 6.6 | 20.9 |
The protein in the table (above) is placed in a buffer solution with a pH of 8.0.
What is the correct net charge on the SL protein at pH of 8.0
Protein Migration Direction Based on Isoelectric Point
Click to show Protein Migration Direction Based on Isoelectric Point example problem
500d
Isoelectric Point Problem
A mixture of two proteins are to be separated by isoelectric focusing.
| Protein Name |
Isoelectric Point (pI) |
Molecular Weight |
|---|---|---|
| Sucrase (Suc) | 6.5 | 51.0 |
| Leghemoglobin (Leg) | 4.7 | 16.0 |
Both protein samples are placed into a gel with a constant pH of 7.5. The gel is then placed into an electric field.
In which direction will each protein in the table migrate at pH 7.5?
Estimating Protein Molecular Weight from SDS-PAGE Migration
Click to show Estimating Protein Molecular Weight from SDS-PAGE Migration example problem
16b5
Gel Migration Problem
In this task, data from an SDS-PAGE experiment, where proteins are separated based on molecular weight, is provided. The gel results table below shows some standard proteins with known molecular weights and one unknown protein.
| Protein Name | Molecular Weight (kDa) |
Migration Distance (cm) |
|---|---|---|
| Cytochrome c (Cyt) | 13.0 | 3.45 |
| Myoglobin (Myo) | 18.0 | 3.19 |
| Succinate Ligase (SL) | 20.9 | 3.07 |
| Elastase II (Ela) | 26.5 | 2.88 |
| Pepsin (Pep) | 34.5 | 2.67 |
| Aldolase (Aldo) | 47.5 | 2.41 |
| DNA Ligase (Lig) | 60.0 | 2.22 |
| Unknown | ? | 2.99 |
Estimate the molecular weight of the unknown protein by comparing its gel migration distance with those of the standards.