CellSig3 - 13 Flashcards Preview

Y2S2 - Cell Signalling > CellSig3 - 13 > Flashcards

Flashcards in CellSig3 - 13 Deck (12):
1

What does endogenous refer to?

Naturally occurring genes, RNA and proteins in cells or organisms

2

What does exogenous refer to?

Experimental manipulation of genes, RNA or proteins added to cells or organisms

3

What sort of systems allow real-time measurement?

Electrophysiology, GFP-tags, biosensors

4

What sort of systems require fixing before measurement?

Immunohistochemistry and biochemistry

5

List three direct labelling techniques

Radioactive, photo/chemically reactive, GFP

6

List the main indirect labelling technique

Immunological - a/b labelling or enzymes etc

7

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

8

Outline immunocytochemistry

Epitope tag incorporated into protein of interest, primary a/b attaches to this tag, then marker-coupled secondaries attach to primary, amplifying response

9

What protein-protein interaction methods can be used?

Co-immunoprecipitation, GST pull-downs, yeast two-hybrid, FRET

10

What does FRET stand for?

Fluorescence resonance energy transfer

11

What in vitro methos are there for studying transcriptional regulation?

Reporter constructs (lacZ, CAT, luciferase), RNAse protection

12

What whole genome approaches are there to studying transcriptional regulation?

Microarrays