A fusion protein can be synthesized to carry a specific tag, which is useful in identifying the proteins in various experiments. Fusion proteins with the FLAG epitope bind to specific anti-FLAG antibodies and carry this extra sequence: Asp-Tyr-Lys-Asp-Asp- Asp-Asp-Lys. a. Draw the structure of the FLAG epitope at pH 7.0. b. As you have drawn it, what is the approximate net charge of the FLAG epitope at pH 7.0? c. How would the structure and net charge of the FLAG epitope change if placed into a pH 12.5 solution?

A fusion protein can be synthesized to carry a specific tag, which is useful in identifying the proteins in various experiments. Fusion proteins with the FLAG epitope bind to specific anti-FLAG antibodies and carry this extra sequence: Asp-Tyr-Lys-Asp-Asp- Asp-Asp-Lys. a. Draw the structure of the FLAG epitope at pH 7.0. b. As you have drawn it, what is the approximate net charge of the FLAG epitope at pH 7.0? c. How would the structure and net charge of the FLAG epitope change if placed into a pH 12.5 solution?

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

See similar textbooks

Similar questions

1) Using the diagram below, sketch in the pattern of bands you would expect to see after digesting the DNA of the TT, Tt and tt genotypes of the TAS2R38 gene. Use the Base Pair Standards on the left of the diagram as a reference in drawing the positions of the bands. Base Pair TASTER GENO TYPE Standards (Base Pairs) TT Tt tt 500 400 300 200 100 50
If a student uses a monoclonal antibody to detect a protein using a direct Western-blotting protocol after running an SDS PAGE, the student can detect a 50kd protein. However, if the student tries to use this antibody to immunoprecipitate the 50 kd protein and then Western -blot the immunoprecipitated samples, the student found that there is no band at all at 50kd region in the Western blot. How do you explain these results based on what you know about protein structure and antigen-antibody recognition? (Hint: Western blot analysis explores an SDS-containing buffer, whereas the immunoprecipitation buffer uses a NP40-containing buffer to maintain proteins in their native form).
True or False: An IgM monomer can bind as many as 10 epitopes?
4) Researcher #1 and #2 in your lab each conducted a protein purification. They present you with the following data. The protein-of-interest is indicated by the red arrowhead, and the numbers on top indicate the purification step. Researcher #1 1 2 3 4 SDS-PAGE (Coomassie Stained) Researcher #2 1 2 3 4 5 6 Immunoblot a) Which researcher has more clearly demonstrated purification of the protein-of-interest? Justify your answer. b) What type of chromatography step did researcher #1 most likely conduct in going from step 2➜ 3 based on the gel? Justify your answer.
Give typing answer with explanation and conclusion What is the transcription product of the DNA templa GCTAGCGATGAC-5'? OA) CAGTAGCGATCG OB) CGAUCGCUACUG OC) CGATCGCTACUG OD) CGUTCGCUTCUG
In a-c below, will the residue on the right-hand side increase or decrease the pKa of the residue on the left-hand side? Explain.
In the human immune system, so-called B cells canmake more than a billion different types of antibodymolecules that protect us from infection. However, ourgenomes have only three genes that encode the polypeptides found in antibodies. What experiments could youperform to determine what kind of combinatorial eventsoccur at the DNA level (V-D-J joining) and RNA level(alternative splicing) for any of these genes?
Monoclonal antibodies are produced by immunizing a mouse with a particular antigen, then removing its spleen. Individual B cells producing mouse antibodies specific for the antigen are isolated from the spleen, then fused with cancer cells. Each of the resulting hybrid cells can be cloned, which means it can be cultured indefinitely in the laboratory. The resulting cell lines produce and secrete antibodies that recognize the antigen to which the mouse was immunized. These antibodies are called monoclonal antibodies. Some monoclonal antibodies are used for passive immunization. They are effective, but only in the immediate term. Antibodies that are produced by one’s own immune system can last up to about six months in the bloodstream, but those delivered in passive immunization often last for less than a week. Why the difference?
How many diff erent heavy chain variable regions can theoretically be generated by somatic recombination in humans (ignore junctional flexibility)? If each of these heavy chains could combine with any of the ~2000 κ light chains, how many different immunoglobulins could be produced?
How can a single nucleotide insertion in the beta globin gene generate a phenotypic change in the ability of the red blood cell to transport oxygen?
A cell is infected with an HIV mutant that results in very little virus being released into the medium that is used to grow the cultured cells. Upon electron microscopy it is noted that the membrane of the infected cell is idenitcal in appearance to a cell that has not been infected. In addition there is relatively little viral RNA in the cells and relatively few proviruses in the cellular genome when compared to a non-mutant HIV infection. Which of the following proteins is most likely mutated? Multiple Choice gag reverse transcriptase vif vpu
. You need to prepare 500 μL of a 250 µg/mL bovine gamma globulin solution, for one of your protein standards in the Bradford assay. The available bovine gamma globulin in the lab is in the form of a 2 mg/mL stock solution, and it can be diluted with deionized water. Describe or show below how you would create your desired 250 µg/mL solution.