User:Maria Briscione/Notebook/Chem 571/2011/09/14

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Objective

Description

A 6mL of a 9.97µg/mL solution (1:1000 dilution of the BSA that comes with restriction enzymes) was created as a working stock solution. Standard solutions of BSA (1mL for each solution) in water of 7.97, 5.98, 3.99, 2.99, 1.99, 1.00 μg/mL were created from the working stock solution. 800µL protein sample (standards or unknown)
and 200µL Bradford reagent (Bio-Rad Protein Assay; purchased Bio-Rad Laboratories, Inc.) were mixed together. Absorbance of standards and unknowns was measured using the UV-vis, and then further analyzed at 595nm. The unknown protein sample was assumed to be at a concentration of 1,000 µg/mL and with this assumption diluted first to a working solution of 10 µg/mL and then further to 2 µg/mL. The absorbance of a blank containing 200 µL of Bradford reagent and 800 µL of distilled water was analyzed. A protein blank consisting of 200 µL of protein and 800 µL.

Data

Image:9-14.jpg

Notes


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