Difference between revisions of "Creating 20.109(S17):Scan slides to identify FKBP12 binders (Day5)"

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==Introduction==
 
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In the previous laboratory session you screened SMM slides using your purified FKBP12 protein.  To identify which of the small molecules you screened bind FKBP12, your slides will be evaluated using a Genepix microarray scanner.  The scanner will measure the fluorescence signal emitted from each spot on the slide where a ligand was printed.  Considering that neither the ligands nor FKBP12 protein is fluorescent, from where does this signal arise?
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[[Image:Sp17 20.109 M1D5 SMM scan.png|thumb|300px|right|]]A positive control was included on each slide.  You may remember that 4 x 48 spots were printed with rapamycin.  Rapamycin is a macrolide compound used to suppress the immune system in organ transplant recipients.  For our purposes, it is a compound known to bind FKBP12.  In the image to the right, the bright red spots show where FKBP12 bound to the rapamycin at these locations.

Revision as of 17:04, 31 January 2017

20.109(S17): Laboratory Fundamentals of Biological Engineering

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Schedule Spring 2017        Announcements        Assignments        Homework        Communication
       1. High-throughput ligand screening        2. Gene expression engineering        3. Biomaterials engineering              

Introduction

In the previous laboratory session you screened SMM slides using your purified FKBP12 protein. To identify which of the small molecules you screened bind FKBP12, your slides will be evaluated using a Genepix microarray scanner. The scanner will measure the fluorescence signal emitted from each spot on the slide where a ligand was printed. Considering that neither the ligands nor FKBP12 protein is fluorescent, from where does this signal arise?

Sp17 20.109 M1D5 SMM scan.png
A positive control was included on each slide. You may remember that 4 x 48 spots were printed with rapamycin. Rapamycin is a macrolide compound used to suppress the immune system in organ transplant recipients. For our purposes, it is a compound known to bind FKBP12. In the image to the right, the bright red spots show where FKBP12 bound to the rapamycin at these locations.