Difference between revisions of "20.109(S18):Confirm ligand binding using differential scanning fluorimetry assay (Day6)"

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(→‎Part 2: Prepare samples for DSF assay)
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==Reagents==
 
==Reagents==
 +
*FKBP12, Abcam
 +
*DSF dye, Thermo Fischer
 +
*ligands, Chembridge
  
 
==Navigation links==
 
==Navigation links==
 
Next day: [[20.109(S18):Complete data analysis (Day7) |Complete data analysis]]<br>
 
Next day: [[20.109(S18):Complete data analysis (Day7) |Complete data analysis]]<br>
 
Previous day: [[20.109(S18):Test protein activity using peptidyl-prolyl cis-trans isomerase assay (Day5) |Test protein activity using peptidyl-prolyl cis-trans isomerase assay]]<br>
 
Previous day: [[20.109(S18):Test protein activity using peptidyl-prolyl cis-trans isomerase assay (Day5) |Test protein activity using peptidyl-prolyl cis-trans isomerase assay]]<br>

Revision as of 22:13, 3 February 2018

20.109(S18): Laboratory Fundamentals of Biological Engineering

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Spring 2018 schedule        FYI        Assignments        Homework        Class data        Communication
       1. Assessing ligand binding        2. Measuring gene expression        3. Engineering biomaterials              


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

  1. 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

  • FKBP12, Abcam
  • DSF dye, Thermo Fischer
  • ligands, Chembridge

Navigation links

Next day: Complete data analysis

Previous day: Test protein activity using peptidyl-prolyl cis-trans isomerase assay
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