BIO314H5 Chapter Notes -Antigen, Nitrocellulose, Luminol
Document Summary
Lab 8 development of immunoblots (western blots) gel has to be transferred to nitrocellulose membrane, this is called protein blotting or western blotting transfer technique is electroblotting since uses electricity: membrane with proteins on it = blot to further investigate, blot is probed with antibody blot is first incubated with blocking agent (i. e. bsa or milk) which binds to all available sites on membrane and blocks them to minimize non specific binding of antibody then incubate with primary antibody followed by secondary antibody which detects the primary interaction of secondary with primary can be visualized on blot one of most common detection methods is to use an enzyme linked secondary antibody following incubation with secondary antibody, the blot is incubated with enzyme substrate solution enzyme converts colourless substrate into insoluble coloured product that is ppt onto membrane an ideal system for blotting is the combination of 4cn and hrp.
