Biology 2382B Lecture Notes - Lecture 3: Primary And Secondary Antibodies, Fluorescence, Antibody
24 Oct 2018
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Isolation and analysis of cell organelles and molecules. Readings: chapter 1: pg 13, chapter 3: p. 105-108; 112-113, chapter 4: p. 161-163, chapter 7: p. 291. Labeling live cells with fluorescent antibodies or stains: antibodies made against specific cell surface proteins can be linked to fluorophores, membrane permeable fluorescent dyes can be used to label intracellular structures (i. e. hoechst stain binds dna in nucleus) Cells with bound antibodies or that have taken up the dyes can now be sorted and counted (note: fluorophore-linked secondary antibodies could also be used in combination with unlabeled primary antibodies) Wbc"s express certain proteins that are transmembrane and stick out on the extracellular side. Example: thy1. 2, cd3: antibody specifically recognize these proteins, we can conjugate these antibodies with flurophore or use secondary antibodies, these are for cell surface proteins. Lets say we want to make sure they are nucleiated proteins.
