Difference between revisions of "User:Moira M. Esson/Notebook/CHEM-571/2013/11/19"

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(AuNP solution preparation)
(AuNP solution preparation)
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==AuNP solution preparation==
 
==AuNP solution preparation==
*The general protocol for the preparation of AuNPs described on [[User  
+
*The general protocol for the preparation of AuNPs described on [[User:Moira M. Esson/Notebook/CHEM-571/2013/08/28|08/28/2013]] was followed.
 +
*20 AuNP solutions were prepared with BSA and Invertase proteins.
 
<br>
 
<br>
 +
Table 1. Gold Standard Solution preparation
 +
<br>
 +
{| {{table}}
 +
| align="center" style="background:#f0f0f0;"|'''Molecular weight Au (g/mol)'''
 +
| align="center" style="background:#f0f0f0;"|'''393.83'''
 +
|-
 +
| mass Au(g)||0.0354
 +
|-
 +
| volume stock(L)||0.025
 +
|-
 +
| Concentration (M)||0.00359546
 +
|}
 +
<br>
 +
Table 2. Preparation of BSA stock solution
 +
<br>
 +
{| {{table}}
 +
| align="center" style="background:#f0f0f0;"|'''Molecular weight (g/mol)'''
 +
| align="center" style="background:#f0f0f0;"|'''66000'''
 +
|-
 +
| mass (g)||0.0432
 +
|-
 +
| volume stock (L)||0.03
 +
|-
 +
| Concentration (M)||2.18182E-05
 +
|}
 +
<br>
 +
Table 3. Preparation of Invertase Stock solution
 +
<br>
 +
{| {{table}}
 +
| align="center" style="background:#f0f0f0;"|'''Preparation Invertase Stock'''
 +
| align="center" style="background:#f0f0f0;"|''''''
 +
|-
 +
| Molecular weight (g/mol)||135000
 +
|-
 +
| mass invertase (g)||0.1015
 +
|-
 +
| volume stock (L)||0.025
 +
|-
 +
| concentration (M)||3.00741E-05
 +
|}
 +
<br>
 +
Table 4. Preparation of BSA-AuNP solutions
 +
<br>
 +
{| {{table}}
 +
| align="center" style="background:#f0f0f0;"|'''Ratio'''
 +
| align="center" style="background:#f0f0f0;"|'''Amount of gold added(mL)'''
 +
| align="center" style="background:#f0f0f0;"|'''concentration BSA (M)'''
 +
| align="center" style="background:#f0f0f0;"|'''volume BSA (mL)'''
 +
| align="center" style="background:#f0f0f0;"|'''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
 +
|}
 +
<br>
 +
Table 5. Preparation of Invertase-AuNP solutions
 +
<br>
 +
{| {{table}}
 +
| align="center" style="background:#f0f0f0;"|'''Ratio'''
 +
| align="center" style="background:#f0f0f0;"|'''Amount gold added (mL)'''
 +
| align="center" style="background:#f0f0f0;"|'''Concentration Invertase (M)'''
 +
| align="center" style="background:#f0f0f0;"|'''Volume Invertase (mL)'''
 +
| align="center" style="background:#f0f0f0;"|'''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
 +
|}
 +
<br>
 +
==Notes==
 +
*All prepared protein-AuNP were colorless. However, the invertase-AuNP solutions prepared had a slightly cloudy appearance.
 +
 
==AA spectroscopy==
 
==AA spectroscopy==
 
<br>
 
<br>

Revision as of 10:59, 19 November 2013

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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

Molecular weight Au (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

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


  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.