User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/11/07

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Project name

Bradford Assay

Conduct a bradford assay on the collected fraction of ADA made on 2012/11/06 and on 2012/09/26.

A stock solution of 1mg/mL BSA was prepared by adding 10mg of BSA into a 10mL volumetric flask and then filling it with autoclaved water. The actual concentration of the BSA stock was 1.02mg/mL as the actual weighed out BSA added was 10.2mg BSA.
In order to determine the concentration of the ADA collected in the fracs, standard solutions had to be made and run through the UV-Vis first. Seven standard solutions were prepared with 1X Bradford Reagent, varying amounts of BSA stock, and autoclaved water. These solutions were analyzed in the UV-Vis from 500-800 nm.

Volume 1X Bradford Reagent Added [mL]	Volume BSA Stock Added [mL]	Volume Water Added [mL]	Conc. BSA [mg/mL]
1.4	0	0.03	0
1.4	0.005	0.025	0.003566434
1.4	0.01	0.02	0.007132867
1.4	0.015	0.015	0.010699301
1.4	0.02	0.01	0.014265734
1.4	0.025	0.005	0.017832168
1.4	0.03	0	0.021398601

The absorbance values at 595 nm of each solution were graphed versus BSA concentration in mg/mL to produce a calibration curve.

When these samples were originally run, they produced absorbances over 1 which resulted in us needing to dilute the solution further. As a result, all solutions were diluted by a half to 715μL of original solution with an additional 715μL of water.