User:Moira M. Esson/Notebook/CHEM-571/2014/02/04

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(Preparation of Hemoglobin:Magnetite Solution)
(Preparation of BSA:Magnetite solutions)
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==Preparation of BSA:Magnetite solutions==
==Preparation of BSA:Magnetite solutions==
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*The  
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**The general protocol for the preparation of protein:magnetite:gold solutions in the oven described on [[User:Moira M. Esson/Notebook/CHEM-571/2014/01/28|01/28/2014]] was followed. However, gold was not added to the reaction mixture (these steps were not included).
*The KNO<sub>3</sub> and KOH stock solutions in Table 6 and Table 7, respectively, were used in this synthesis.
*The KNO<sub>3</sub> and KOH stock solutions in Table 6 and Table 7, respectively, were used in this synthesis.
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*The FeSO<sub>4</sub>*7H<sub>2</sub>O stock solution in Table 2 was used.
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*The FeSO<sub>4</sub>*7H<sub>2</sub>O stock solution in Table 2 was used.
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<bR>
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Table 9. BSA Stock solution preparation
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{| {{table}}
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| align="center" style="background:#f0f0f0;"|'''Preparation of BSA stock'''
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| align="center" style="background:#f0f0f0;"|''''''
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|-
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| Molecular weight (g/mol)||66463
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|-
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| mass (g)||0.0167
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|-
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| volume stock (L)||0.01
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|-
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| Concentration (M)||2.51268E-05
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|}
<br>
<br>
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Table 10. BSA:Magnetite solution preparation
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<br>
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{| {{table}}
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| align="center" style="background:#f0f0f0;"|'''Ratio'''
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| align="center" style="background:#f0f0f0;"|'''concentration BSA (M)'''
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| align="center" style="background:#f0f0f0;"|'''volume of BSA added (mL)'''
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| align="center" style="background:#f0f0f0;"|'''amount of FeSO4 added (mL)'''
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| align="center" style="background:#f0f0f0;"|'''amount of KOH added (mL)'''
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| align="center" style="background:#f0f0f0;"|'''amount of KNO3 added (mL)'''
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| align="center" style="background:#f0f0f0;"|'''volume water added (mL)'''
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|-
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| 10000||0.000003||0.596973054||0.5||0.500624554||2.5||0.902402392
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|-
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| 13000||2.30769E-06||0.459210041||0.5||0.500624554||2.5||1.040165405
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|} 
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<br>
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==AA sample preparation==
==AA sample preparation==
*All samples prepared on [[User:Moira M. Esson/Notebook/CHEM-571/2014/01/28|01/28/2014]] were centrifuged at 2000 rpm for 30 minutes.
*All samples prepared on [[User:Moira M. Esson/Notebook/CHEM-571/2014/01/28|01/28/2014]] were centrifuged at 2000 rpm for 30 minutes.

Revision as of 15:26, 4 February 2014

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Objectives

  1. Prepare a Hemoglobin:Magnetite solution following the general protocol described on 01/29/2014
  2. Prepare a hemoglobin:Magnetite solution of the same ratio following the general protocol described on 01/28/2014 for comparison
  3. Prepare BSA:Magnetite solutions of the same ratio using the general method of protein-Au preparation in the oven described on 01/28/2014.
  4. Centrifuge BSA:Au:Magnetite soluions prepared on 01/28/2014. This will be used for AA analysis tomorrow. The supernatent will be collected, dried, and analyzed using DSC, FTIR and X-ray.


Preparation of Hemoglobin:Magnetite Solution

Method A. Schlenk Line Preparation

  • The general protocol described on 01/29/2014 was followed.

Table 1. Preparation of Hemoglobin stock solution

Preparation of hemoglobin Stock solution '
Molecular weight (g/mol)64500
mass hemglobin (g)0.1
volume solution (L)0.1
Concentration (M)1.55039E-05


Table 2. Preparation of FeSO4*7H2O stock solution

Preparation of FeSO4 stock '
Molecular Weight (g/mol)278.01
mass(g)0.83403
volume stock (L)0.01
Concentration (M)0.3


Table 3. Preparation of KOH stock solution

Preparation of KOH stock '
Molecular Weight (g/mol)56.11
mass(g)0.5606
volume stock (L)0.01
Concentration (M)0.999108893


Table 4. Preparation of KNO3 stock solution


Preparation of KNO3 '
Molecular Weight (g/mol)101.1
mass(g)0.5604
volume stock (L)0.01
Concentration (M)0.5


  • After beginning the synthesis, it was discovered that this stock was mis-prepared, leading to the concentration listed above. The concentration was supposed to be 0.8M. A new synthesis of hemoglobin:magnetite using the Schlenk line method will need to be performed.


Table 5. Preparation of Hemoglobin:Magnetite solution

Ratio concentration Hemoglobin (M) volume of Hemoglobin added (mL) amount of FeSO4 added(mL) amount of KOH added (mL) amount of KNO3 added (mL) volume Water added (mL)
130002.30769E-060.7442307690.50.5004459512.50.75532328


  • The ratio is two orders of magnitude larger than the ratio when gold is used for prep due to the fact this ratio refers to the concentration of FeSO4 in solution (which is 0.03M). As such, the ratio was altered to make similar concentration comparisons (with this order of magnitude it is comparing to 0.003M which is similar to the 0.0025M gold in the previous solutions)


Method B. Oven Preparation

  • The general protocol for the preparation of protein:magnetite:gold solutions in the oven described on 01/28/2014 was followed. However, gold was not added to the reaction mixture (these steps were not included).


Table 6. Preparation of KNO3 stock solution

Preparation of KNO3 '
Molecular Weight (g/mol)101.1
mass(g)4.044
volume stock (L)0.05
Concentration (M)0.8


Table 7. Preparation of KOH stock solution

Preparation of KOH stock '
Molecular Weight (g/mol)56.11
mass(g)0.5604
volume stock (L)0.01
Concentration (M)0.998752451


Table 8. Preparation of Hemoglobin:Magnetite solution using the oven

Ratio concentration Hemoglobin (M) volume of Hemoglobin added (mL) amount of FeSO4 added(mL) amount of KOH added (mL) amount of KNO3 added (mL) volume Water added (mL)
100000.0000030.96750.50.5006245542.50.531875446
130002.30769E-060.7442307690.50.5006245542.50.755144677


  • The FeSO4*7H2O stock solution described in Table 2 was used for this synthesis


Preparation of BSA:Magnetite solutions

    • The general protocol for the preparation of protein:magnetite:gold solutions in the oven described on 01/28/2014 was followed. However, gold was not added to the reaction mixture (these steps were not included).
  • The KNO3 and KOH stock solutions in Table 6 and Table 7, respectively, were used in this synthesis.
  • The FeSO4*7H2O stock solution in Table 2 was used.


Table 9. BSA Stock solution preparation

Preparation of BSA stock '
Molecular weight (g/mol)66463
mass (g)0.0167
volume stock (L)0.01
Concentration (M)2.51268E-05


Table 10. BSA:Magnetite solution preparation

Ratio concentration BSA (M) volume of BSA added (mL) amount of FeSO4 added (mL) amount of KOH added (mL) amount of KNO3 added (mL) volume water added (mL)
100000.0000030.5969730540.50.5006245542.50.902402392
130002.30769E-060.4592100410.50.5006245542.51.040165405


AA sample preparation

  • All samples prepared on 01/28/2014 were centrifuged at 2000 rpm for 30 minutes.


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