7.02 and 7.021 require simultaneous registration. Application of experimental techniques in biochemistry, microbiology, and cell biology. Emphasizes integrating factual knowledge with understanding the design of experiments and data analysis to prepare the students for research projects. Instruction and practice in written communication provided.
The venom of the honeybee, Apis mellifera, contains numerous enzymes, volatiles, and other components. Allergic individuals develop anti-venom antibodies which may cause hives, anaphylactic shock, and death. In this exercise students detect concentrations of bee venom on nitrocellulose membranes following incubation of membranes with anti-venom antibodies, enzyme-conjugated secondary antibodies, and a colorimetric substrate. The intensity of the colored substrate deposited on the membrane is proportional to the original concentrations of bee venom spotted on the membrane. Students are introduced to micropipetting and to the concept of detection limits while learning about allergy and immunoassays.
This is a collection of tried-and-true technique descriptions used in teaching postgraduate students in the Department of Molecular & Cell Biology at UCT.
Students perform Western blots with polyacrylamide gel electrophoresis of detergent extracts of SV40-transformed cell lines grown in monolayer culture. The gels are blotted onto nitrocellulose and treated with a monoclonal antibody against SV40 T antigen or against the tumor suppressor p53. Blots are then treated with a secondary antibody (anti-mouse IgG conjugated to horseradish peroxidase), and exposed to revealing chemicals and x-ray film. I give one application: induction of p53 by ultraviolet light.
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