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Discuss the limitations of differential centrifugation as a means of isolating subcellular components.
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- What is differential centrifugation? Explain how it can be used to separate cellular organelles.The following questions refer to membrane protein isolation A) What is the difficulty when working with membrane proteins and extracting them from the membrane? B) Why do you need stabilization of membrane proteins? What reagents are used for stabilization? C) What methods are used to determine if your extracted membrane protein (GPCR) is functional? D) What methods are used for the enrichment of surface proteins? E) What 2 methods are used to analyze and identify surface proteins after extraction?What could be an example of in vitro experiment to identify cytoskeleton accessory proteins?
- discuss some diseases associated with flawed cytoskeletons – Kartagener’s syndrome, blistering syndromes.Using two centrifuges (A and B), with maximum speeds of 1500 rpm (r = 8 cm) and 3000 rpm (r = 8 cm), respectively, construct a separation scheme for separating three subcellular structures with the following xg requirements: PAT – 150 xg - SAT – 120,000 xg НАТ - 650 хgDescribe and connect five different outcomes that could occur due to the loss of FMRP function in the cell in the figure provided.
- Describe the two phenomena that give rise to the diversity of adhesion molecules such as cadherins. What additional phenomenon gives rise to the diversity of integrins?Cancer stem cells (CSCs) are cancer cells (found within tumors or hematological cancers) that possess characteristics associated with normal stem cells, specifically the ability to give rise to all cell types found in a particular cancer sample. There are many biomedical engineering based approaches to detect CSCs. 1) What is the importance and advanatge of detecting CSCs? 2) What kind of systems have been developed to detect CSCs? Describe by giving examples.Protein transfer to a membrane from a developed electrophoretic gel can be performed under which of the following conditions?
- Using two centrifuges (A and B), with maximumspeeds of 1500 rpm (r = 8 cm) and 3000 rpm(r = 8 cm), respectively, construct a separationscheme for separating three subcellular structureswith the following xg requirements:• PAT – 150 xg• SAT – 120,000 xg• HAT – 650 xgFor this exercise, an electron micrograph of secretory cells of the anterior pituitary is being used (Sandborn, E.B. (1970) Cells and Tissues by Light and Electron Microscopy, New York, New York: Academic Press, Inc.). The image originally measured 18.7 cm wide and 18.5 cm high with a magnification of 5200x. Given the width of the original micrograph at 18.7cm, what is the true width of unmagnified image? Show calculations.Outline an experimental approach to measure Amyloid Beta that microglia produces in vitro. What are some of the problems you might encounter?