Class_1_highlights - the protein and a secondary antibody...

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Class 1 highlights: What is a gel filtration column? On what basis does it separate proteins? ‐A gel filtration column is filled with small porous beads. Large proteins do not interact with beads and flow through quickly. Small proteins get hung up in beads. ‐You often use this to separate protein complexes from one another‐ so they can be run under mild conditions so that protein‐protein interactions are maintained. How does a Western blot work? 1. how are the proteins separated?‐ SDS‐PAGE‐ make sure you understand how this works. 2. How are the proteins detected?‐ using a primary antibody directed against
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Unformatted text preview: the protein, and a secondary antibody that binds the first one, and also has an acitivity that allows it to be “seen” 3. Make sure you understand on what basis proteins are separated on SDS‐ PAGE gels‐ where do smaller and larger proteins run?, and generally how antibodies work. Look back and remind yourself of the difference between a monoclonal and polyclonal antibody. Which is likely to have more binding sites in any particular protein? A monoclonal will only recognize one epitope in a protein, but in a polyclonal there could be many different antibodies that recognize different epitopes....
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