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WorksheetsProtein Purification and Field TripQuiz
Total questions: 29
Worksheet time: 16mins
Hydrophobic amino acids are ones which _______ water.
are attracted to
are repelled by
react with
melt in
What “feature” did the Hydrophobic Interaction Chromatography (HIC) beads have to bind to protein?
Nickel ions
HIS
Methyl (-CH3)
Polyethylene Glycol
In HIC, 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 Hydrophobic Interaction Chromatography, how was the protein flipped inside out?
Genetic engineering
The protein was mixed with PEG and centrifuged
A high salt solution was added
A solution free of salt was added.
In HIC, 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 HIC, how did adding a low salt solution help with removing the GFP from the beads
It didn't help.
It reacted with the proteins and turned purple
It caused the GFP to flip back to their original shape
It added HIS-tag to the end of the protein
In "HIS-tag" protein purification, what is "HIS"?
Histidine - an amino acid
Histamine - a chemical produced by the body during an immune reaction
Hydrophobic Interaction Systems
Highly Immunologic Serum
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, how was a HIS-tag added to the protein?
Genetic engineering
The protein was mixed with PEG and centrifuged
A high salt solution was added
A solution free of salt was added
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
What is the purpose of BCA?
It binds to beads and causes protein to fall off
It turns purple in the presence of protein so protein can be measured
It is a reference protein with known concentrations
It measures the intensity of the purple color
What does a spectrophotometer do?
It causes proteins to fall off of the beads
It turns purple in the presence of proteins
It measures the intensity of the purple color
It is a reference protein of known concentration
What is the purpose of BSA in protein purification?
It causes the proteins to fall off of the beads
It turns purple in the presence of amino acids
It measures the intensity of the purple color
It is a reference protein of known concentration
What is the purpose of measuring the intensity of the purple color of BSA mixed with BCA if we already know the concentration of BSA? CHECK ALL WHICH APPLY
You never know what could happen
It allows us to make a standard curve
It allows us to compare the known concentration of BSA with the A562 of our samples
It causes the protein to fall off of the beads
What is a possible conclusion from this graph?
Oranges cost more per pound than apples
Oranges and plums are statistically the same price
Oranges and applies are statistically the same price
Oranges cost twice as much as apples on average
The standard curve for this graph shows that the data is good because
It is straight
It has a slope of less than 1
It has an R2 close to 1
The standard curve for this graph shows that the data is NOT good
In protein gel electrophoresis, what has to be done to the protein samples before loading the protein gel?
Denatured using heat
Denatured using a chemical called beta-mercaptoethanol (BME)
Uniformly covered with a negative charge using SDS
All of the above
Why was it important to do denature a protein and uniformly cover it with a negative charge before loading proteins into the gel?
Different proteins have different shapes which affects how they would run in the gel
Different proteins have different charges which affects how they would run in the gel
Proteins must have the right shape in order to do their job but we don’t want them doing their job in a gel
A and B
All of the above
How large is GFP?
200 kDa
40 kDa
27 kDa
10 kDa
Which lane shows the purest GFP sample?
M
I
FT
E
What is your evidence that lane E is the purest GFP?
It has the darkest blue band
It only has one band
It was loaded last
It isn't the purest - lane I is the purest
What was the overall purpose of the field trip lab?
See how much pizza you could eat
Miss a day of school
Learn how antibodies are purified and how they can be used to detect disease
Learn how antigens could be purified and how they can be used to detect disease.
What was the purpose of adding BSA to the nitrocelluose membrane (the paper which was sticky for all types of proteins)?
They help to find the antigen
They help to bind to IgY
They turn TMB blue
They fill in the empty spaces to “block” the rest of the membrane
What was the purpose of the primary antibody (the ones from the egg)?
They help to find the antigen
They help to bind to IgY
They turn TMB blue
They fill in the empty spaces to “block” the rest of the membrane
What was the purpose of the secondary antibody (the ones bound to horseradish peroxidase)?
They help to find the antigen
They help to bind to IgY
They turn TMB blue
They fill in the empty spaces to “block” the rest of the membrane
What was the purpose of the horseradish peroxidase?
They help to find the antigen
They help to bind to IgY
They turn TMB blue
They fill in the empty spaces to “block” the rest of the membrane
What are two “real world” applications of this immune system technology? (2 pts)
