User:Moira M. Esson/Notebook/CHEM-571/2013/11/19: Difference between revisions
From OpenWetWare
(fix raw html notebook nav) |
|||
(9 intermediate revisions by 2 users not shown) | |||
Line 2: | Line 2: | ||
|- | |- | ||
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | ||
|style="background-color: #F2F2F2" align="center"| | |style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
|- | |- | ||
| colspan="2"| | | colspan="2"| | ||
Line 10: | Line 10: | ||
#Analyze AuNP-protein solutions prepared on [[User:Moira M. Esson/Notebook/CHEM-571/2013/10/30|10/30/2013]] using AA spectroscopy. | #Analyze AuNP-protein solutions prepared on [[User:Moira M. Esson/Notebook/CHEM-571/2013/10/30|10/30/2013]] using AA spectroscopy. | ||
<br> | <br> | ||
==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> | |||
Table 1. Gold Standard Solution preparation | |||
<br> | |||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"|''' Preparation of Gold Stock Solution''' | |||
| align="center" style="background:#f0f0f0;"|''''''''' | |||
|- | |||
|Molecular Weight (g/mol)||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;"|'''Preparation of BSA stock solution''' | |||
| align="center" style="background:#f0f0f0;"|'''''' | |||
|- | |||
|Molecular weight (g/mol)||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== | |||
<br> | |||
'''General Protocol''' | |||
#Transfer 3mL of sample from test tubes into a 10mL falcon tube. | |||
#Centrifuge 3mL protein-AuNP solutions for 30 minutes at 4000rpm. Be sure that the centrifuge is balanced. | |||
#Prepare 5 standard Au solutions (10μg/mL, 20μg/mL, 30μg/mL, 40μg/mL, 50μg/mL) following protocol described on [[User:Moira M. Esson/Notebook/CHEM-571/2013/09/10|09/10/2013]]. | |||
#Run samples on AA. | |||
<br> | |||
Table 6. Standard Au Solution preparation from 1000μg/mL AA Au gold standard solution | |||
<br> | |||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"|'''Concentration(μg/mL)''' | |||
| align="center" style="background:#f0f0f0;"|'''Volume of stock Au solution(mL)''' | |||
| align="center" style="background:#f0f0f0;"|'''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 | |||
|} | |||
<br> | |||
Figure 1. Calibration curve of gold standard solutions | |||
<br> | |||
[[Image:Calibration Curve of Gold Standards zem11192013 .png]] | |||
<br> | |||
Figure 2. Atomic Absorbance Spectra of Myoglobin:AuNP solutions | |||
<br> | |||
[[Image:Atomic Absorbance Spectra of Myoglobin AuNP solutions zem11192013.png]] | |||
<br> | |||
Figure 3. Atomic Absorbance Spectra of Lysozyem:AuNP solutions | |||
<br> | |||
[[Image:Atomic Absorbance Spectra of Lysozyme AuNP solutions zem 11192013 .png]] | |||
<br> | |||
*The only solution that was colored was the 30 ratio. All other solutions appeared colorless. | |||
Figure 4. Absorbance vs. Concentration plot of Myoglobin:AuNP solutions | |||
<br> | |||
[[Image:Absorbance vs. Concentration plot of Myoglobin-AuNP solutions zem11192013.png]] | |||
<br> | |||
Figure 5. Absorbance vs. Concentration plot of AuNP-Lysozyme Solutions | |||
<br> | |||
[[Image:Absorbance vs. Concentration plot of Lysozyme-AuNP solutions zem11192013.png]] | |||
Latest revision as of 23:35, 26 September 2017
Project name | Main project page Previous entry Next entry | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Objectives
AuNP solution preparation
Notes
AA spectroscopy
Figure 4. Absorbance vs. Concentration plot of Myoglobin:AuNP solutions
|