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Sept. 20 |
Laboratory Safety and Procedures |
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Check out the abbreviated instructions for this week. |
Cupboard assignment and Equipment |
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Calibration of Pipetman |
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Experiment 1 |
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Determination of Protein Concentration |
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Bradford Method |
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UV Absorbance assay |
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Sept. 27 |
Experiment 2 |
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Check out the abbreviated instructions for this week. |
Determination of the Activity and Specific Activity of ß-Galactosidase |
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Prepare Dialysis Tubing |
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Lab Report Covering Experiments 1 and 2 - due October 6. |
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Oct. 4 |
Experiment 7 |
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Now we are going to isolate some b-galactosidase. See the abbreviated instructions. |
Isolation of HisĄTag Overexpressed ß-Galactosidase: Use of BugBuster and Benzonase |
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Read: |
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Experiment 3: Isolation and Purification of ß-Galactosidase by the Classical Method: Ammonium Sulfate Precipitation |
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Experiment 8 |
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Using the affinity chromatography method, the ß-Galactosidase will be purified. Here are the abbreviated directions. |
Purification of HisĄTag Overexpressed ß-Galactosidase: Immobilized Metal Affinity Chromatography |
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Assay Fractions for both Protein and Enzyme Activity |
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Read: |
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Experiment 4: Purification of ß-Galactosidase: Ion Exchange Chromatography |
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Experiment 5: Purification of ß-Galactosidase: Gel Filtration Chromatography |
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Experiment 6: Purification of ß-Galactosidase: Affinity Chromatography |
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Oct. 11 |
No Lab |
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Oct. 18 |
Experiment 9 |
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Follow directions inthe manual |
SDS Gel Electrophoresis |
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Oct. 25 |
No Experiments Except to Take Pictures of the Gels |
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Lab Report Covering Experiments 7, 8 and 9 - due November 3. |
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Nov. 1; Nov. 8; Nov. 15 |
Experiments 10, 11, 12 , and 13 or experiments suggested by the individual groups based on the following experiments or on other literature sources.
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Experiment 10 |
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Determination of the Stability of Protein Structure |
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and/or |
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Experiment 11 |
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Cofactor Requirements and the Effect of pH on ß-Galactosidase Activity |
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and/or |
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Experiment 12 |
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The Qunatitation and Chemical Modification of the Cysteine Resudes in ß-Galactosidase |
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and/or |
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Experiment 13 |
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Enzyme Kinetics and the Binding of Ligands to ß-Galactosidase Determination of Km and Ki |
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Nov. 29 |
Organize Cupboards |
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Lab Report Covering Experiments 10, 11, 12, and 13 and/or other experiments - due December 1. |
See the course description for Chem/Biol/BIMO 321