Biochemistry: Concepts and Connections
1st Edition
ISBN: 9780321839923
Author: Dean R. Appling, Spencer J. Anthony-Cahill, Christopher K. Mathews
Publisher: PEARSON
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Textbook Question
Chapter 6, Problem 14P
A protein gives a single band on SDS get electrophoresis, as shown in lanes 1 and 2 below. There is little, if any, effect from adding
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Chapter 6 Solutions
Biochemistry: Concepts and Connections
Ch. 6 - Prob. 1PCh. 6 - Bovine pancreatic trypsin inhibitor (BPTI; Figure...Ch. 6 - A schematic structure of the subunit of...Ch. 6 - In the protein adenylate kinase, the C-terminal...Ch. 6 - Give two reasons to explain why a proline residue...Ch. 6 - Consider a small protein containing 101 amino acid...Ch. 6 - a. Based on a more conservative answer to Problem...Ch. 6 - The following sequence is part of a globular...Ch. 6 - a. A protein is found to be a tetramer of...Ch. 6 - Under physiological conditions, the protein...
Ch. 6 - Theoretical and experimental measurements show...Ch. 6 - The peptide hormone vasopressin is used in the...Ch. 6 - A protein gives under conditions of buffer...Ch. 6 - A protein gives a single band on SDS get...Ch. 6 - It has been postulated that the normal...Ch. 6 - Prob. 16PCh. 6 - Do you expect a Pro Gly mutation in a...Ch. 6 - Rank the following in terms of predicted rates...
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- Calculate these for 1 ug of plasmid DNA in a final total volume of 20 ul. All enzymes being used here are at a concentration of 10 U/µL. For double digests, give the appropriate volume for each enzyme and decide which buffer to use. (HINT: look up double digest conditions on the NEB website.) You should always calculate all volumes in advance to ensure the correct working concentrations and so that you can prepare the digests as efficiently as possible. It is a good idea to check off each component as it is added to the microfuge tube. Complete the table below, including the volume of DNA sample(s) you will need for 1 pg of the DNA (from two different methods in Experiment #3) based on the concentration(s) you determined from the OD260 value, which you will restriction digest with each of the restriction enzymes singly and in double digests in Part A. For example, the volume of DNA sample that contains 1 µg DNA for DNA sample from Part A, Experiment 3: OD260 = 0.024, dilution 500x…arrow_forwardin isolating ribosomes from a yield sample, describe the ideal type of centrifugation for this separation technique based on the following:*Type of Centrifugation* Type of fraction *Give 2 advantages of using this type of centrifuge.*Give 2 disadvantages of using this type of centrifuge.arrow_forwardElectrophoretic Analysis of DNA via Agarose gel Electrophoresis determine the voltage used (how to determine the voltage? Hint: check on volt/cm).arrow_forward
- In an in vitro motility assay, a newly discovered motor protein is found to transport an attached bead along a microtubule away from the end where fluorescent labeled tubulin dimers assembled. Which of the following are true? Select all that apply The motor protein is transporting the bead towards the minus end The motor protein is transporting the bead towards the plus end The motor protein belongs to the dynein family The motor protein belongs to the kinesin family Oarrow_forwardTransferring of the protein bands from the SDS-PAGE gel to the PDVF membrane can be troublesome and sometimes incomplete. What are some reasons (at least 3) why the transfer step wouldn’t work?arrow_forwardIn your own words, describe how we will use hydrophobic interaction chromatography to purify GFP from the multitude of other proteins in a bacterial cell. Be sure to include the purpose of each of the wash solutions in the process (Equilibration, Binding, Wash, and Elution Buffers).arrow_forward
- Explain how gel filtration chromatography works. What type of gel will you used when the protein size is 2500 Da? Explain.arrow_forwardGiven this, if you used 6g of vitamin Z powder to make 20 ml of solution, what is the % concentration of this solution? (I gave the image since I don't know if that info is needed to solve this question.)It also gives a follow-up, if you can help here too: You work in a lab as a summer student. One of your tasks is to make sure that there is enough cell culture medium containing antibiotics to grow bacteria. One day you realize that there is only 5 ml of 10% Antibiotic stock solution in the freezer. You decide to use it all to prepare the working culture medium with 0.01% antibiotic. In the lab there is plenty of growth medium without antibiotics. (Note: dilution in medium is like dilution in water). You remember the equation to make dilutions of stock solutions. You usually use this formula to calculate the required volume of a stock solution, but you realize it can apply here as well, even though the unknown is the final volume. So, you make that dilution. Given that each bacterial…arrow_forwardProtein analysis by gel electrophoresis a). Using the gel image provided to calculate the electrophoretic relative mobility as a ratio of the distance of protein migration to the distance of the tracking dye migration (See Appendix B). If your dye front is not visible, measure the mobility relative to the bottom edge of the resolving gel. Include a labeled print-out of your gel image with your report. b). Plot log MW of the protein markers and commercial myoglobin vs. mobility. Determine the molecular weight of myoglobin obtained after the final purification step (Sample E) from the equation of the line. Submit a copy of your graph along with the gel image. c). Compare your sample E and commercial myoglobin with the ladder. Summary your results. order of sample (left to right) is : A B C D blank E blank Commercial Myoglobin Molecular Ladderarrow_forward
- You overexpressed (asked bacteria to make a lot of) an 80 kDa protein carrying a 6X-His tag and purified it on a nickel column. You collect frac ons from your purifica on and analyze them using SDSPAGE. The results are shown in image attached. a) Interpret the results from the gel. What is wrong? (see image attached) b) Identify 2 possible reasons why this problem is occurring, and for each reason, suggest an experimental modifica on you can do to troubleshoot.arrow_forwardHIC chromatography does not yield 100% pure GFP. What other types of cellular proteins would most likely be found in the GFP preparation?arrow_forwardBesides agarosc gel electrophoresis, there is also SDS-PAGE for separation of macromolccules. Differentiate both these methods.arrow_forward
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