bis_104_midterm_exam_i_key2_fall_07 - Page 5

bis_104_midterm_exam_i_key2_fall_07 - Page 5 - cells. OR:...

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Page 4 Name ______________________________ It has been hypothesized that the proliferation of these breast cancer cells is beyond normal control because they over-express the erbB2 receptor. 10. Describe one experiment to test the hypothesis that the cancer cells have more erbB2 receptors in their plasma membranes than normal cells. List all of your key steps in chronological order. (4) Direct immunocytochemistry approach: a. obtain pure populations of both normal and breast cancer cells. b. Prepare or purchase a fluorochrome labeled antibody specific for human erbB2 protein. c. treat both live cell batches with equal concentrations of the antibody; allow time to bind; wash away any unbound antibody. d. observe cells by fluorescence microscopy and compare the intensity of PM fluorescence; normal vs cancer
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Unformatted text preview: cells. OR: Biochemical approach: a. Start with pure populations of normal and cancer cells. b. Isolate and purify PMs from both ell types. c. do a total protein extraction from the membrane preps. d. separate extracted proteins by SDS-PAGE; do western blot and probe for erbB2 with labeled antibody. e. compare density of labeled protein bands on blot. Describe the results you would expect to obtain if your hypothesis were proven to be correct. (4) Cells taken from tumor tissue would show more intense fluorescence when compared to cells from normal tissue. OR: PM protein extracts from cancer cells would yield higher concentration of erbB2 protein per cell as revealed by western blot of normal vs cancer cell PMs. Demo...
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