User:Moira M. Esson/Notebook/CHEM-571/2013/11/19

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Objectives

  1. Prepare AuNP-protein solutions for biomineralization
  2. Analyze AuNP-protein solutions prepared on 10/30/2013 using AA spectroscopy.


AuNP solution preparation

  • The general protocol for the preparation of AuNPs described on 08/28/2013 was followed.
  • 20 AuNP solutions were prepared with BSA and Invertase proteins.


Table 1. Gold Standard Solution preparation

Preparation of Gold Stock Solution ''''
Molecular Weight (g/mol) 393.83
mass Au(g) 0.0354
volume stock(L) 0.025
Concentration (M) 0.00359546


Table 2. Preparation of BSA stock solution

Preparation of BSA stock solution '
Molecular weight (g/mol) 66000
mass (g) 0.0432
volume stock (L) 0.03
Concentration (M) 2.18182E-05


Table 3. Preparation of Invertase Stock solution

Preparation Invertase Stock '
Molecular weight (g/mol) 135000
mass invertase (g) 0.1015
volume stock (L) 0.025
concentration (M) 3.00741E-05


Table 4. Preparation of BSA-AuNP solutions

Ratio Amount of gold added(mL) concentration BSA (M) volume BSA (mL) volume Water added (mL)
105 0.347660664 2.38095E-06 0.545634921 4.106704416
110 0.347660664 2.27273E-06 0.520833333 4.131506003
115 0.347660664 2.17391E-06 0.498188406 4.15415093
120 0.347660664 2.08333E-06 0.477430556 4.174908781
125 0.347660664 0.000002 0.458333333 4.194006003
130 0.347660664 1.92308E-06 0.440705128 4.211634208
135 0.347660664 1.85185E-06 0.424382716 4.22795662
140 0.347660664 1.78571E-06 0.40922619 4.243113146
145 0.347660664 1.72414E-06 0.395114943 4.257224394
150 0.347660664 1.66667E-06 0.381944444 4.270394892


Table 5. Preparation of Invertase-AuNP solutions

Ratio Amount gold added (mL) Concentration Invertase (M) Volume Invertase (mL) Volume water added (mL)
30 0.347660664 8.33333E-06 1.38546798 3.266871356
50 0.347660664 0.000005 0.831280788 3.821058548
70 0.347660664 3.57143E-06 0.593771992 4.058567345
90 0.347660664 2.77778E-06 0.46182266 4.190516676
110 0.347660664 2.27273E-06 0.377854904 4.274484432
130 0.347660664 1.92308E-06 0.31972338 4.332615956
150 0.347660664 1.66667E-06 0.277093596 4.37524574
170 0.347660664 1.47059E-06 0.244494349 4.407844987
190 0.347660664 1.31579E-06 0.218758102 4.433581234
210 0.347660664 1.19048E-06 0.197923997 4.454415339


Notes

  • All prepared protein-AuNP were colorless. However, the invertase-AuNP solutions prepared had a slightly cloudy appearance.

AA spectroscopy


General Protocol

  1. Transfer 3mL of sample from test tubes into a 10mL falcon tube.
  2. Centrifuge 3mL protein-AuNP solutions for 30 minutes at 4000rpm. Be sure that the centrifuge is balanced.
  3. Prepare 5 standard Au solutions (10μg/mL, 20μg/mL, 30μg/mL, 40μg/mL, 50μg/mL) following protocol described on 09/10/2013.
  4. Run samples on AA.


Table 1. Standard Au Solution preparation from 1000μg/mL AA Au gold standard solution

Concentration(μg/mL) Volume of stock Au solution(mL) Volume Deionized water added(mL)
10 0.1 9.9
20 0.2 9.8
30 0.3 9.7
40 0.4 9.6
50 0.5 9.5