Biochemistry
Biochemistry
8th Edition
ISBN: 9781285429106
Author: Campbell, Mary K., FARRELL, Shawn O.
Publisher: Cengage Learning,
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Chapter 5, Problem 3RE

RECALL What is meant by “salting out”? How does it work?

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You were tasked with purifying an untagged transcription factor (molecular weight 65,000Da, isoelectric point unknown) from a contaminant Protein A (molecular weight 50,000 Da, isoelectric point 5.0). You were also instructed to use the protein for crystallization studies after purification.i) In the initial purification step, which type of chromatography should you attempt? Explain your choice and specify the requirements of the buffer solution you should use. ii) Analyzing the proteins recovered from step i) using SDS-PAGE, you still observed a faint band of Protein A in addition to the transcription factor band. Given the limited time available for further purification, you must choose ONE appropriate chromatography method to maximize the chances of separating the two proteins as well as your crystallization studies. Provide a detailed explanation of your selection and the techniques/strategies you would employ to achieve this.
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Chapter 5 Solutions

Biochemistry

Ch. 5 - RECALL What is the basis for the separation of...Ch. 5 - RECALL What is the order of elution of proteins on...Ch. 5 - RECALL What are two ways that a compound can be...Ch. 5 - Prob. 14RECh. 5 - RECALL Why do most people elute bound proteins...Ch. 5 - RECALL What are two types of compounds that make...Ch. 5 - RECALL Draw an example of a compound that would...Ch. 5 - RECALL How can gel-filtration chromatography be...Ch. 5 - REFLECT AND APPLY Sephadex G-75 has an exclusion...Ch. 5 - Prob. 20RECh. 5 - RECALL What is the main difference between reverse...Ch. 5 - RECALL How does HPLC differ from ion-exchange...Ch. 5 - REFLECT AND APPLY Design an experiment to purify...Ch. 5 - REFLECT AND APPLY Referring to Question 23, how...Ch. 5 - Prob. 25RECh. 5 - REFLECT AND APPLY You wish to separate and purify...Ch. 5 - REFLECT AND APPLY An amino acid mixture consisting...Ch. 5 - REFLECT AND APPLY An amino acid mixture consisting...Ch. 5 - REFLECT AND APPLY In reverse-phase HPLC, the...Ch. 5 - REFLECT AND APPLY Gel-filtration chromatography is...Ch. 5 - RECALL What physical parameters of a protein...Ch. 5 - RECALL What types of compounds make up the gels...Ch. 5 - RECALL Of the two principal polymers used in...Ch. 5 - RECALL What types of macromolecules are usually...Ch. 5 - RECALL If you had a mixture of proteins with...Ch. 5 - RECALL What does SDSPAGE stand for? What is the...Ch. 5 - RECALL How does the addition of sodium...Ch. 5 - RECALL Why is the order of separation based on...Ch. 5 - RECALL The accompanying figure is from an...Ch. 5 - Prob. 40RECh. 5 - Prob. 41RECh. 5 - Prob. 42RECh. 5 - Prob. 43RECh. 5 - REFLECT AND APPLY What would happen during an...Ch. 5 - REFLECT AND APPLY A sample of an unknown peptide...Ch. 5 - REFLECT AND APPLY A sample of a peptide of unknown...Ch. 5 - REFLECT AND APPLY You are in the process of...Ch. 5 - REFLECT AND APPLY You are in the process of...Ch. 5 - Prob. 49RECh. 5 - Prob. 50RECh. 5 - Prob. 51RECh. 5 - Prob. 52RECh. 5 - Prob. 53RECh. 5 - Prob. 54RECh. 5 - RECALL What is the basis for the technique called...Ch. 5 - Prob. 56RECh. 5 - Prob. 57RECh. 5 - RECALL What are the main procedures involved in a...Ch. 5 - RECALL Where did western blot get its name?Ch. 5 - Prob. 60RECh. 5 - Prob. 61RECh. 5 - Prob. 62RECh. 5 - Prob. 63RECh. 5 - Prob. 64RECh. 5 - Prob. 65RE
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