bis_104_pq_16a_ans_08 - Cells from normal individuals show...

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BIS 104 Practice Question #16 a answers 1. Plasma membrane lacks receptors for LDLs. Experiment: Prepare or obtain a fluorescently-labeled or ferritin-labeled antibody specific for LDL receptor protein. Obtain fibroblasts from the skin of both normal individuals and FH- individuals and grow in cell culture. Apply labeled antibodies to both cell preps; allow time to bind; wash away any unbound antibody. Examine (fluor microscopy or EM, depending on label) and compare labeling of normal vs FH- cells. Expected results:
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Unformatted text preview: Cells from normal individuals show positive plasma membrane labeling, whereas the FH-fibroblast lack labeling. (alternative experiment: Isolate plasma membranes from both cell types by differential centrifugation; verify purity by TEM of samples. Separate PM proteins by SDS-PAGE; transblot (Western) separated proteins and probe both samples with a labeled antibody specific for LDL receptor protein. Results as above: PM extracts from normal cells are positive; FH- cells are negative...
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