WorksheetsLIFE2087 SFAP - Protein purification
Total questions: 13
Worksheet time: 7mins
Ion-Exchange chromatography:
separate and analyze compounds that can be vaporized without decomposition
Also known as Liquid-Solid Chromatography
Materials used are either cation or anion resins
Separation based on partition coefficient between 2 immiscible liquids
What type of chromatography is depicted by the illustration?
Gel Filtration Chromatography
Column Chromatography
HPLC
Thin Layer Chromatography
Which of the following methods could be used to check the molecular weight of your purified protein?
SDS-PAGE
Mass Spectrometry
Analytical SEC
All of above
To elute target proteins from an affinity chromatography matrix, which of the following conditions would be the most appropriate?
Low salt concentrations
High salt concentrations
Adding a soluble ligand which competes with the affinity tagged protein for binding to the column
Just keep washing buffer through the column, isocratic elution
What is the starting point for selection of a suitable IEX matrix for purification of a recombinant protein?
Prediction of isoelectric point (pI) from the amino acid sequence
Test protein binding to an IEX matrix at a range of pHs and salt concentrations
Test protein binding to a selection of anion and cation exchange matrices
Pass your sample through a preparative column and elute with a salt gradient
You find that your protein sample loses activity during storage. What can you do about this?
Add an additional purification step
Use a protease inhibitor during purification steps
Perform each step as quickly as possible, in a cold-room
All of the above
Hydrophobic amino acids are ones which _______ water.
are attracted to
are repelled by
react with
melt in
What “feature” did the His-Tag beads have to bind to protein?
Nickel ions
HIS
Methyl (-CH3)
Polyethylene Glycol (PEG)
In HIS-tag protein purification, what had to be done to GFP before adding the GFP to the beads?
A HIS tag was added
The protein was precipitated with PEG
The protein was flipped inside out
The protein was extracted from the egg
In HIS-tag protein purification, what had to be done in order to get the GFP to fall off the beads?
Add imidazole
Add an even higher concentration of PEG
Add a high salt solution
Add a solution free of salt
In HIS-tag protein purification, how did adding imidazole help to remove the GFP from the beads?
The beads flipped inside out
The protein flipped inside out
The beads dissolve when mixed with imidazole
Nickel prefers imidazole over HIS-tags
