ANAT 262 Lecture Notes - Lecture 3: Green Fluorescent Protein, Polyclonal Antibodies, Direct Fluorescent Antibody

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Recall: use of fluorescent microscope to visualize proteins by shining light on the sample (short wavelength) which has fluorescent dyes. The sample will emit light at longer wavelength. By setting different blocks, we can see different fluorescent proteins at the same time. E. g. microtubules (tubulin) glowing green and er glowing red. To visualize this molecule we added antibodies against the protein of interest. We preferred to work with thin tissue sample. Tissues will have to be sectioned most of the time, if not we could not see very much at the microscope. Labeling molecules in cells: immunofluorescence, immunogold, microinjection, green fluorescent protein. The idea is that you add an antibody which is conjugated to a fluorescent dye or not (direct and indirect). Direct is the simplest method where you have an antibody conjugated with fluorescent dye, and you add it to cells. The antibody will bind to antigen and emit fluorescent light.

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