User:Jacob Esenther/Notebook/Chem 571/2014/09/10

Biomaterials Design Lab

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Objective Explore the use of Bradford Analysis with Lysozyme Description Bradford Analysis Prepared 50 mL of a standard saline solution (0.9 wt-% NaCl). Prepared 50 mL of a 50 mM Tris (not Tris-HCl) 50 mM NaCl solution. Prepared a stock solution of BSA that is roughly 5 mg in 5 mL of saline. Calculated your actual solution concentration. Using a quartz cuvette, recorded UV-VIS spectra between 200 nm and 800 nm. Prepared serial dilutions of 1μg/mL, 2μg/mL, 2.5μg/mL, 5μg/mL, 8μg/mL, 10μg/mL, 15μg/mL and 20μg/mL Added 20μL of the serial dilutions to a 1.5mL centrifuge tube Added 200μL of the Bio-Rad Protein Assay reagent. Use 1:4 concentrate diluted with water. Added 780μL of Tris/NaCl buffer to make the final volume 1mL. Obtained a UV-VIS spectrum. Blanks of Tris/NaCl buffer and 200 μLBradford reagent + 800μL buffer were also run Data Bradford Assay with Lysozyme Bradford Absorbance vs. Concentration Bradford Absorbance vs. Concentration (outliers removed) BSA and Lysozyme Stock Solutions for Bradford Corrected Bradford-BSA Absorbance Notes No notes to report