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problem set 2.5 - – another divalent cation b You find...

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15. Challenge: ELISA assays are commonly used to detect the presence of a molecule, often a protein, in a biological sample. a. Explain how you could use an ELISA assay to detect which monoclonal antibody hybridoma clone is expressing an antibody that is specific to the antigen that was originally injected into the rabbit/mouse. b. The ProteinChip SELDI system was described in class and compared to the ELISA system. Why might the ProteinChip system be considered the “next generation” ELISA system? In other words, what are the advantages of this system compared to conventional ELISAs? 16. Challenge: Figure 9-15 in your text shows the change in calcium levels in a moving leukocyte. a. How could you confirm that this apparent increase in intracellular calcium is due exclusively or in part to calcium and not in total, or in part, to magnesium
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Unformatted text preview: – another divalent cation? b. You find that when you do a similar experiment with nerve cells and add a calcium ionophore to the medium there is a rapid and very large increase in intracellular calcium followed by leakage of the Fura-2 from the cell. What other fluorescent dye discussed in class might be useful to determine if the cell membrane is rupturing due to this increase in intracellular calcium versus an active pump mechanism that is activated by high calcium that, in turn, pumps the Fura-2 out of the cell? c. You find that when you add an ATP inhibitor (a drug that prevents the formation of ATP by blocking mitochondrial oxidative phosphorylation) there is also a big rise in intracellular calcium. But in this case no ionophore is present. What might you presume about the basis underlying this observation?...
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