20.109(S18):Confirm ligand binding using differential scanning fluorimetry assay (Day6)
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Contents
Introduction
Protocols
Part 1: BE Communication Lab workshop
Our communication instructors, Dr. Sean Clarke and Dr. Prerna Bhargava, will join us today for a workshop on writing impactful abstracts and titles.
Part 2: Prepare samples for DSF assay
- Each team will setup triplicate reactions for 5 different conditions:
- Condition 1: control protein (?)
- Condition 2: control protein AND control substrate (?)
- Condition 3: no protein
- Condition 4: FKBP12
- Condition 5: FKBP12 AND rapamycin
- Condition 6: FKBP12 AND ligand #1
- Condition 7: FKBP12 AND ligand #2
| Reagent (stock concentration) | Final concentration of stock reagent in reaction | Volume of stock reagent in reaction |
|---|---|---|
| FKBP12 (1 mg/mL) | 1.2 μg/mL | |
| DMSO (1%) | 0.2% | |
| rapamycin (50 μM) | 10 μM | |
| ligand (200 μM) | 40 μM | |
| dye (5X) | 1X | |
| PBS (1X) | add for a total of 10 μL per reaction |
Reagents
Next day: Complete data analysis
