Lecture 7 Other Cell Biology Techniques Flashcards
What is a yeast-2-hybrid
2 domains of a transcription factor are separated and each attach to a different protein
Used to study: if 2 proteins fused to domains bind each other
What is FACS
used to separate cells based on special properties
What is Cell Fractionation
separate part of a cell by centrifugation
used to find out where in a cell a protein is
What is differential centrifugation
used to study how fast components spin to the bottom
What does the low speed pellet contain
nucleus
What does the low speed supernatant contain
cytoplasm
What does the high speed pellet contain
most organelles except ER fragments and fragment plasma membrane pieces
What does the high speed supernatant contain
ribosomes and proteins
What is equilibrium centrifugation
spin until components stop moving in density gradient
separates components based on density
What is a pulse chase
used to study where something moves in a cell
can be monitored by microscopy or cell fractionation
What is the CMC. what happens when detergent is above/below the CMC?
CMC is the concentration at which it will solubilize micelles?
If the detergent is above the CMC then the membrane proteins will be solubilized in micelles
If the detergent is below the CMC then the membrane proteins will be solubilized individually
Ionic vs non-ionic detergents
ionic- strong, denature
non-ionic- weak, solubilize
What is DNA Ligase
seals nicks and attaches DNA
What are vectors
a carrier for the DNA of interest
Kinds of vectors?
viruses and plasmids
plasmids are the most common
