Lab 7 Flashcards
1
Q
Steps of protein purification
A
1) Cell lysis
2) Clarification (removal of debris)
3) Extraction (denature protein)
4) Separation/isolation
5) Detection/analysis
2
Q
Two types of cell lysis
A
Gentle (osmotic, enzymatic)
Vigourous (sonication)
3
Q
3 main types of protein separation
A
1 - Separation by size
2 - Ion exchange
3 - Affinity tag
4
Q
Spectrophotometer
A
Measures optical density
5
Q
Epitope
A
Short amino acid sequence on a protein that binds to a ligand - added to the protein of interest
6
Q
In this lab what do the cells have?
A
Cells have IPTG -
compound that inhibits the lac repressor; induces protein expression
7
Q
What tag are we using in this lab?
A
Purify mNeonGreen using a His-Tag added to C terminal
This binds to Nickel ions - use a nickel resin
8
Q
A
