Biology 2290F/G Lecture Notes - Lecture 3: Hoechst Stain, Cd90, Differential Centrifugation

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Labelling live cells with fluorescent antibodies or stains. Lecture 3: isolation and analysis of cell organelles & molecules. Cells with bound antibodies or that have taken up the dyes can now be sorted and counted. Say we want to characterize a certain type of white blood cells, which express a certain type of proteins. They"re transmembrane proteins, with a bit that sticks out on the outside. They have these things called cd3 and thy1 2. Antibodies made against specific cell surface proteins can be linked to fluorophores. Membrane permeable fluorescent dyes can be used to label intracellular structures (e. g. hoechst stain binds dna in nucleus) We have an antibody that recognizes thy1, and another one that recognizes cd3. Antibodies can be conjugated to fluorophores, where they can recognizes proteins on the outside of the cell. We can add membrane permeable fluorescent dyes that can be used to label intracellular structures.

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