Physiology 3140A Chapter Notes - Chapter 3: Exonuclease, Reverse Transcriptase, Embryonic Stem Cell
5 Oct 2017
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Incubate in antibody retrieval solution to remove aldehyde groups: wash sections with pbs between steps, blocking = incubate sections in hydrogen peroxide to prevent nonspecific antigen binding (peroxidases) to glass slide. Incubate with primary antibodies to detect protein of interest. Immunocytochemistry: culture media removed from cell layer growing in culture dish, cells do not need to be embedded in wax (not tissue, western blot. Isolate protein on ice remove growth medium: lyse cells by adding detergent, collect protein sample in eppendorf tube. Inhibitors can be added in vitro to identify how agents affect a ligand-gated channel based on if they produce inward or outward current. Imprecise matching allows targeting of many mrnas: targeting can lead to degradation or inhibition of translation, argonautes and their proteins are found in plants, animals, fungi, and some bacteria. Video: ddrnai vector enters nucleus shrna expressed, passes through to cytoplasm, shrna is constitutively expressed stable knock-down.
