2. What factors might change your decision?7. Imagine you're purifying a protein with a pl of 6.5 that has a 6-Histidine tag from E. coli lysate. You're planning to use Ni-NTA resin (nickel resin)↓which has a high affinity for the 6-His tag, to purify your protein from the rest of the lysate.1. Which pH ranges would you expect a protein with a pl of 6.5 to be stable in? (Remember: pl is isoelectric point)2. What pH range is recommended by Ni-NTA resin manufactures for purification?3. What pH do you think would work best for your purification process and which buffering agent(s) would work well for that pH?8. A lab mate prepared a 50 mM Tris buffer for you, and told you that it was pH 8. But the next day, you took the buffer out of the refrigerator and

Ni-NTA agarose is an affinity chromatography matrix that is used for the purification of recombinant…

Answered: 7. Imagine you're purifying a protein…

Biochemistry

9th Edition

ISBN:9781319114671

Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.

Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.

Chapter1: Biochemistry: An Evolving Science

Section: Chapter Questions

Problem 1P

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1. What is the isoelectric point (pI) of lysine which has pKa values of 2.1 for the α carboxyl group, 9.7 for the α amino group and 10.5 for the side chain amino group? 2. Which of the following is most likely to be found on the exterior of a protein? A) Pro B) Trp C) Ser D) Glu 3. The type of reaction that forms a peptide bond is A) Elimination B) Hydrolysis C) Nucleophilic substitution D) Condensation
1. Suppose Biuret test is conducted to a solution of RNA. Will it give a positive result or not? Explain your answer. ____________________________________________________________________ ____________________________________________________________________ 2. Why is Molisch’s test used for the determination of presence of pentose in the hydrolysate? What other test could be used for this? ____________________________________________________________________ ____________________________________________________________________
2. A ligand binds more tightly to the folded state (N) of a protein than to the unfolded state (U). Show that the ligand stabilizes the protein and calculate by how much (ΔΔGfold = ?). Please help me find out deltadelta G fold.
2 a. You are trying to purify protein C from a mixture of proteins noted in the above Table. If you had only one type of column to choose from, which one would allow you to purify protein C with the least number of contaminants? Size exclusion column Ion exchange column Affinity chromatography using glucose as the bait Affinity chromatography using NAD as the bait Please explain why you chose the column above based upon the properties of the column AND the proteins in the Table.
3 Common Databases on Protein Study (Peptide Sequence to Structural Analysis).A tool provided before 2020 and after 2020? Your task is to find : 1) Disadvantage/limitation of the mentioned database Common Databases on Protein Study (Peptide Sequence to Structural Analysis).A tool provided before 2020 and after 2020?
1. What is the name of the nucleotide shown here?2. Identify all pointed structures (A to H)3. Which structure causes the acidity of the nucleotide? Choose theletter of your answer.
1.Draw these phosphorylated structures as they would be connected in a polinucleotide (e.g.RNA) in the order A-B. / Show how they combine to form the polynuleotide (i.e. only the end product). Show at any one of these structures where the glycosidic bond occurs 2.Sanger sequencing revealed the sequence of an oligonucleotide to be: d-AGATGCCTGACT. Draw a diagram of the gel banding pattern post capillary electrophoresis i.e. where on the gel would the fragments feature
1.a.If this molecule was the side chain of an amino acid, in a protein, what tertiary structure stabilizers could be present? b.Make a key and use color to highlight what part of you structure can be stabilized in each manner?
2. Would you expect an instrinsically disordered protein to contain a higher proportion of hydrophilic or hydrophobic residues? Explain your reasoning.
4. What is the region/point where AA is predominantly present as a (-2)-charged species?5. The effective buffering range for the amino acid in the acidic region is?6. What is the region/point where the solution has a 50:50 percent mixture of the (0) and the (-1) species?
SDS-PAGE gels are used to study changes in protein primary structure. Which of the following statements about SDS-PAGE gels is correct? A. The negatively charged proteins will move towards the negatively charged electrode in the SDS-Page chamber B. A detergent is needed to disrupt protein secondary, tertiary, and quaternary structure before a gel can be run C. No "stains" or other treatments are needed to "see" the protein bands on a gel D. Long polypeptides move a further distance from the loading well in a SDS-PAGE gel than short polypeptides
pls explain why Which of these CANNOT BE achieved using SDS-PAGE?I. Estimate the MW of a protein.II. Confirm the identity of a protein.III. Confirm the presence of quaternary structure in the protein.IV. Give a qualitative assessment of the protein extract purity.V. Determine the shape of the protein.a. II and Vb. III and IVc. I, III and Vd. I, II, and IV Which of the following statements are TRUE about the discontinuous gel in SDSPAGE?I. The sandwich effect occurs because of the lower mobility of the zwitterionic form of glycine compared to the protein band in the stacking gel.II. Because of the larger pores in the resolving gel, proteins move faster in the stacking gel, but decelerate at the boundary of the two gels.III. In the stacking gel, in terms of net negative charge of the species, the order is: zwitterionic glycine > proteins > chloride ion.IV. The higher pH in the resolving gel allows glycine to carry a more negative charge, resulting in higher mobilities compared to…

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