Protein Fluorescence Flashcards
(8 cards)
Why do we use fluorescence
Provides info on molecular environment and on dynamic processes
What is fluorescence
The light emitted by an atom or molecule after a finite duration subsequent to the absorption of electromagnetic energy
Things that can be measured with fluorescence
- temp PH Viscosity Ions Polarity
Instrumentation in fluorescence
Fluorimeters
Microscopes
High throughout plate readers
Intravital imaging
What is the dominant intrinsic fluorophore?
Tryptophan
3 ways to make a GFP fusion construct
Protein x - GFP
GFP - protein y
Protein -GFP - z
Purpose of FRET
Fluorescence resonance energy transfer
Allows the detection of molecule-molecule interactions in the nanometer range with light microscopy
MDCK clues
Madin-Darby canine kidney polarized epithelial cells