BCMB20005 Lecture 7: Lecture 7

• School: University of Melbourne
• Department: Biochemistry and Molecular Biology
• Course: BCMB20005
• Professor: Amber Willems

Document Summary

Activity assays: measure the activity of the protein of interest. Electrophoretic analyses: analyze relative amounts of proteins, including protein of interest: native electrophoresis (agarose or page, denaturing sds-page (enables size determination, 2-d gel electrophoresis. Quantitative protein assays: measure the total concentration of protein , not just protein of interest, e. g. lowry or bradford protein assays. Using uv spectrophotometry to determine the amount or concentration of protein in a sample. *absorbance at 280 nm: due to presence of aromatic amino acid residues within the protein e. g. tyrosine and tryptophan. Tryptophan have 2 other bumbs given by the spectro charateristics of the aromatic group. Using dyes and a standard curve to determine the amount or concentration of protein in a sample. Take portion of protein in solution and react it with reagent that produce color product. Two main groups of assays based on chemistry involved: Binding of protein-specific dyes (reagents), e. g. biuret assay. Each dye binds to different component of the protein.