HTHSCI 1I06 A Lecture Notes - Lecture 5: Polysome, Immunohistochemistry, Protein C
6 Nov 2018
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Produced to help re-fold misfolded proteins in the cytosol. Sugar group is added covalently to n terminus. Required for proper folding and further transport to the. The combination of a mrna + multiple ribosomes. Membrane bound: produce proteins to be released from cell or are destined for the membrane. Immunoblotting: isolate + purify protein sample, separate protein sample through chromatography, wash out non-specific stain + apply antibodies to determine target protein. Immunohistochemistry: prepare tissue slice on slide, apply 1 antibody, wash + apply reporter tagged 2 antibody, apply enzyme link and a chromogen, view colour product with microscope. Enzyme-linked immunosorbent assay (elisa: absorb enzyme linked to ab to a surface, add sample (antigen, add enzyme substrate (colour/light, optically detect signal of enzyme/substrate reaction. Edman degradation: digest sample proteins into smaller fragments, amino terminal is cleaved w/out disturbing other peptide bonds. Each amino acid is identified as it is removed. Mass spectroscopy: measures mass-to-charge ration, enzyme digest, vaporize, ionize, detect.
