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- Immunofluorescence and Fluorescent livecell imaging techniques can both be used to determine protein localisation.List the advantages and disadvantages of using immunofluorescence andfluorescent live cell imagingCompare and contrast the following protein characterization techniques in terms of the principles governing their functions. size exclusion chromatography vs ion-exchange chromatographyYou got an opportunity to join a professor lab who is working in-vivo model and specifically looking at the dysregulation of mitochondria in liver. He asked you to isolate mitochondria from a Rat liver and placed in an assay medium. Based on the knowledge you gain in this course so far, please answer the following questions: a) Which technique will you use to isolate mitochondria? b) What happens to the pH of the medium when the medium is kept anaerobic? c) What happens when O2-saturated saline is added to the mixture?
- The following are words used in animal testing and its alternative methods. Define the following terms - a. Vivisection b. Optogenics c. In vivo d. In vitro cell culture e. In silico computer simulation f. Pharmacokinetics g. Pharmacodynamics h. Tissue culture i. Phototoxicity j. 3D cell culturesA high-throughput assay is being conducted in a 96 well plate to test compounds for anti-bacterial activity. Live bacterial cells are detected using “Live clear", a dye which is initially coloured blue, but turns clear in the presence of live bacterial cells. The control wells are shown below. Penicillin kills bacteria; glycerol does not affect them. Which of the control wells will appear blue at the end of the assay? "Live clear" glycerol penicillin - - + (A1 A2)(A3)A4) bacterial cells B1 (B2 (B3)B4 А4, В1, B2, ВЗ, B4 A2, A4, B2, B3, B4 А4, B2, ВЗ, В4 A1, АЗ, В1, В2, вЗ, В4 + + |3) Define gel electrophoresis, including its theory and application. Describe the steps of running gel electrophoresis using the following image. More detailed reading: https://www.sciencedirect.com/topics/medicine-and-dentistry/agar-gel-electrophoresis POWER SUPPLY CATHODE ELECTROPHORETIC BUFFER ANODE WELL SAMPLE AGAROSE GEL POWER SUPPLY CATHODE ANOCE HIGH MOLECULAR WEIGHT SPECIES LOW MOLECULAR WEIGHT ANALYTES
- Among the techniques below, which method is used to study protein-protein interaction in vivo? Group of answer choices co-IP TAP Y2H All of the abovein isolating ribosomes from a yield sample, describe the ideal type of centrifugation for this separation technique based on the following:*Main type of centrifuge *Subtype centrifuge * Speed range *Operational temperature range*Type of CentrifugationWhat are the functions of the following steps in the purification and isolation of proteins: Freeze-thawing the cells • Cell press • Addition of lysozyme and DNase • Sonication • Centrifugation
- Proteins secreted by bacteria can be used as biomarkers for diagnostics. Give a concise account of how you can use a 2D gel electrophoresis combined with mass spectrometry to develop a diagnostic for E. coli infection. Discuss how you could extend this technique to develop a diagnostic for COVID-19 based on protein electrophoresis?A protein gives a single band on SDS gel electrophoresis, as shown in lanes 1 and 2 below. There is little, if any, effect from adding β-mercaptoethanol(BME) to the sample; if anything, the protein runs a little bit slower. When treated with the proteolytic enzyme thrombin and electrophoresis in the absence of BME, the protein migrates a bit more rapidly (lane 3). But if BME is present, two much more rapidly migrating bands are found (lane 4). Explain these results in terms of a model for the protein.Correctly identify the steps and outcomes in the process of screening for a nutritional mutant of a bacterium. Drag the appropriate labels to their respective targets.