BIOL 301 Lecture Notes - Lecture 8: Fusion Protein, Ubiquitin, Myc
Document Summary
After purifying your poi, what can you do with it: study/confirm ppi in a complex, e. g. Sds page + western blot: assess biochemical function in vitro, e. g. Milder, weakly denature: mild = use for protein in cytoplasm, no need to break nuclear membrane, non-polar, break membranes, triton x-100. Mild, doesn"t denature proteins, too mild for extracting: results: you get a soup of proteins, jellybean analogy membrane proteins (use only for soluble?, purify/separate/isolate: many different methods separate by different properties of protein. Affinity chromatography: tag the protein with a prey, put. But you need to tag the protein (go thru cloning/transforms etc. Which can be a hassle or impossible, in which case use a combination of the other co-purifications) Chromatography = purification in a column, with resin (solid agarose beads) Batch = similar to chromatography, except use cups instead of column.
