BCHM 426 Lecture Notes - Lecture 8: Lysis, Bradford Protein Assay, Acrylamide
18 Apr 2017
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Run on fblc on continuous trace to detect proteins. By the intensity, you can tell which fraction has your protein. Put together gel and want to load crude lysate in row 1. Elution fractions are in rows 3, 4, and 5. Look for the size of the band and get an idea of how much protein there is by the intensity of the band. Separating gel for this lab sds will be 15% acrylamide. The taller your spacer is, the more you can fit in the well. The property that the acrylamide concentration affect is how big the pores are. A higher conc of acrylamide, the smaller the pores. That"s what this tells us because it is a highly cross linked, small gel. Success determined by calculation specific activity of the enzyme and protein yield. Couple parameters that tell you about your protein. How much protein you get how pure it is.
