Flashcards in CellSig3 - 13 Deck (12):
What does endogenous refer to?
Naturally occurring genes, RNA and proteins in cells or organisms
What does exogenous refer to?
Experimental manipulation of genes, RNA or proteins added to cells or organisms
What sort of systems allow real-time measurement?
Electrophysiology, GFP-tags, biosensors
What sort of systems require fixing before measurement?
Immunohistochemistry and biochemistry
List three direct labelling techniques
Radioactive, photo/chemically reactive, GFP
List the main indirect labelling technique
Immunological - a/b labelling or enzymes etc
Outline fusion protein incorporation
Gene for protein of interest has DNA encoding epitope/GFP tag inserted, then introduced into cell; natural incorporation leads to purification, visualisation or immunolocalisation
Epitope tag incorporated into protein of interest, primary a/b attaches to this tag, then marker-coupled secondaries attach to primary, amplifying response
What protein-protein interaction methods can be used?
Co-immunoprecipitation, GST pull-downs, yeast two-hybrid, FRET
What does FRET stand for?
Fluorescence resonance energy transfer
What in vitro methos are there for studying transcriptional regulation?
Reporter constructs (lacZ, CAT, luciferase), RNAse protection