BIOC 301 Study Guide - Final Guide: Bradford Protein Assay, Restriction Digest, Enzyme Kinetics
Document Summary
1 after 3 years of graduate studies, you have managed to purify a protein to the point where you are able to crystalise it. At this point, you can assume that the protein is, essentially, pure. You decide to work out how much of the protein you actually have. Using an online tool (which is not down for website maintenance) you determine that the expected extinction co-efficient at 280nm for the protein under your spectrophotometry conditions is 250 00 m -1 , cm -1 . You prepare 100ul of a 1:10 dilution of your protein and do a spec reading at. 2 given the following graph, the formula in the enzyme kinetics documents and that the inhibitor concentration was. 4mm, determine km and ki for this system (substrate, enzyme, inhibitor). Assume plasmid was ligated into the plasmid using the hindii site. Assume all restriction digest sites are at 425 for ease of calculation.
