ANAT 262 Lecture Notes - Lecture 4: Monoclonal Antibody, Fluorescence Microscope, Polyclonal Antibodies

• School: McGill University
• Department: Anatomy & Cell Biology
• Course: ANAT 262
• Professor: John Presley

Document Summary

Lecture 4: light microscopy iii dimanche 24 janvier 2016. We can quantify the amount of fluorescence with digital cameras now. Start with live cells in tissue culture, fix them with formaldehyde to immobilize the proteins and then permeabilize the cell you can then add antibodies (direct vs indirect) Polyclonal antibodies: what you get when you immunized an animal. Monoclonal antibodies: uniform population directed against same antigen. First step is to immunize a mouse, and then you will express large amount of lymphocytes and then you will merge them with myeloma line. You will place one cell per well and screen them one by one. If you find one that is good, you can make a cell line producing antibodies. You cannot produce monoclonal antibodies from lots of species. Gfp: it is possible to attach gfp at n and c-terminus of fusion protein such that they can fold properly.