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

From OpenWetWare
Owwnotebook icon.png Project name <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

Bradford Assay



  • 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.
  • Using linear regression, the equation of the line was obtained.

Bradford assay.png

  • Fractions of ADA, made on 2012/10/03 and 2012/11/06, were obtained.
  • 7.5 μL of each fraction was added to 7.5 μL of autoclaved water and 715 μL of 1X Bradford reagent.
    • Fraction #2, obtained in November, was diluted by half, using the same procedure as listed above for the standard solutions, such that its absorbance was below 1.
Sample Name Volume of ADA [mL] Absorbance [] Conc. ADA [mg/mL] Mass of ADA [mg] Moles of ADA [mol] Molar Absorptivity of ADA [mL/mg cm] Molarity [M]
  • 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.