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When considering anion exchange chromatography, which of the following statements best describes how increasing concentrations of NaCl elute the protein from the column?

Option A: The proteins that are bound to the chromatography resin are exchanged for Cl–.

Option B: The Na+ ions that are bound to the chromatography resin are exchanged for proteins.

Option C: The proteins that are bound to the chromatography resin are exchanged for Na+.

Option D: The Cl– ions that are bound to the chromatography resin are exchanged for proteins.

Given your answer to the question above, which of the following methods would be best suited for a one-step purification of lactate dehydrogenase from a hypothetical mixture of the three proteins?

Option A: anion exchange chromatography using a pH 9 buffer

Option B: anion exchange chromatography using a pH 5 buffer

Option C: anion exchange chromatography using a pH 10.5 buffer

Option D: anion exchange chromatography using a pH 7 buffer

Which of the following combinations of methods would be best suited for the purification of cytochrome c from a hypothetical mixture of the three proteins?

Option A: Gel filtration chromatography to obtain the low molecular weight proteins, followed by cation exchange chromatography at pH 8.

Option B: Cation exchange chromatography at pH 10, followed by elution at high [NaCl], and final purification using gel filtration chromatography.

Option C: Anion exchange chromatography at pH 10, followed by elution at high [NaCl], and final purification using gel filtration chromatography.

Option D: Cation exchange chromatography at pH 7, followed by elution at high [NaCl], and final purification using gel filtration chromatography.

How many amino acids are in the longest form of the human DNA methyltransferase?

Option A: 1511

Option B: 1882

Option C: 1606

Option D 1678

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Related questions

A mixture of three proteins (A,B,C) can be thereetically separated by anion exchange chromotgraphy using cellulose coated with diethylamino-ethyl groups (pka 10.5). In the first step the mixture was applied to the resin at pH 7.5 and washed with the same buffer to elute protein(s) that did not bind. The resin was then washed with a buffer at pH 4.5

Protein A IS A HOMODIMER WITH A MOLECULAR WEIGHT OF 50 kda AND A pi OF 3.0 PROTEIN b IS A HETEROTETRAMER WITH A MOLECULAR WEIGHT OF 100 Kda AND A pi OF 6.0 PROTEIN C IS A MONOMER WITH A MOLECULAR WEIGHT OF 75 Kda AND A pI OF 9.0

WHICH OF THE FOLLOWING STATEMENTS IS INCORRECT?

A. PROTEINS A AND B ARE INITIALLY BOUND TO THE RESIN AT PH 7.5

B. THE RESIN ITSELF WILL REMAIN POSTITIVELY CHARGED OVER THE PH RANGE 3 TO 7.5

C. ONLY PROTEIN A WILL ELUTE WHEN THE RESIN IS WASHED WITH A BUFFER AT PH 4.5

D. PROTEIN C DOES NOT BIND TO THE RESIN AT PH 7.5

1. Which of the following statements is NOT true of the behavior of proteins in SDS
PAGE?
a. The rate of movement of the proteins depends on their charge density
b. The rate of movement of the proteins depends on their shape
c. The rate of movement of the proteins depends on their isoelectric point
d. The rate of movement of the proteins depends on their mass
e. All of the above is true

2. Suppose you ran an ion-exchange chromatography and discovered that three proteins
were all bound by the column. Assuming that this is the only column that you have, what
would be the best strategy to get the samples to elute as distinct peaks?
a. Change the pH of the elution buffer to a new value
b. Apply a gradient of pH
c. Apply a gradient of Salt
d. All of the above
e. b and c

1. Joshua used gel filtration chromatography to purify protein Z from crude homogenate prepared from Arabidopsis plants. He noticed that protein Z appeared to have eluted from the column in two separate peaks. Using a standard curve, he calculated the MW of protein Z eluted in the first peak to be 156 KDa, and the protein Z eluted in the second peak had the apparent MW of 78 KDa. Which one do you think is the correct MW? What does this tell you about the structure of the protein?

2. Read the following paragraph and correct all errors (4.15 points).

An unemployed high school student, Summer, has isolated a protein from a juniper tree with an overall negative charge. He performs ion exchange chromatography using a cationic resin with blue dextran to aid the proteins association with the column. As the proteins come off the column, the spec shows 4 peaks, only the first peak (wash peak) shows activity in an enzyme assay. Since all fractions associated with this peak had activity, they were pooled and total protein assayed. Total protein analysis was assayed using Bicine reagent which interacts with proteins to cause a change in absorption when measured with a spectrophotometer. The results of the total protein calculation showed that there was a modest decrease in total protein which means that there was a large amount of pure protein of his interest present in the sample.

3.You have accidentally mixed a bunch of amino acids (tryptophan, tyrosine, alanine, selenocysteine, proline, arginine) together and once your boss finds out, you will be fired on the spot. Your only hope of saving your job is to separate them before anyone finds out. You decide that the only viable option is to use ion exchange chromatography. Explain, in detail, how you would do this? (4.15 points)

4. Why should you not lose any LDH in your 20k x g spin? How does this explain an increase the fold purification of LDH in your 20k x g sample compared to your crude homogenate? (2 points)

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