BIOL 311 Lecture Notes - Lecture 10: Fluorophore, Gap Junction, Photobleaching

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Frap (fluorescence recovery after photobleaching: measures membrane mobility which proteins move within lipid bilayer, steps: Mobile proteins will redistribute (regain color) laser photobleaches protein. Some gap junctions are only one way and differ in size and acceptance of charges. Fluorescence immunocytochemistry locate cellular molecules (proteins) using fluorochromes and antibodies: requirements, fluorochrome, antigen molecule that elicits an immune response. Need to identify: epitope binding site, antibody. Typically bivalent (able to bind to two antigens) 21-22*: considerations about antibodies, specificity ability of an antibody to bind to an antigen, affinity tightness how well it binds. Making antibodies: polyclonal antibody inject the antigen into animal, presented to b cells. Millions of b cells with different antibodies. 22*: problems, cross reactivity (same antibody binding) Interfere with each other: when animal dies, antibodies die too, monoclonal antibodies, single clone, single antibody, immortal (lasting forever) unlimited source, made by fusing b cells and lymphoma b cells.

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