User:Moira M. Esson/Notebook/CHEM-571/2014/01/29

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(Preparation of BSA-Magnetite solution)
(Preparation of BSA-Magnetite solution)
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#Prepare a 0.8M stock solution of KNO<sub>3</sub> in a 50mL volumetric flask using deionized H<sub>2</sub>O.
#Prepare a 0.8M stock solution of KNO<sub>3</sub> in a 50mL volumetric flask using deionized H<sub>2</sub>O.
#Add the necessary amount of BSA for the desired ratio of BSA to Magnetite to a 10mL round bottom flask.
#Add the necessary amount of BSA for the desired ratio of BSA to Magnetite to a 10mL round bottom flask.
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#
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#Add the appropriate volume of FeSO<sub>4</sub>*7H<sub>2</sub>O to each test tube so that the solution has a concentration of 0.03M FeSO<sub>4</sub>*7H<sub>2</sub>O.
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#Add the appropriate volume of KOH to each test tube so that the solution has a concentration of 0.1M KOH.
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#Add the appropriate volume of KNO<sub>3</sub> to each test tube so that the solution has a concentration of 0.4M KNO<sub>3</sub>.
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Preparation of hemoglobin magnetic fibers at a 130:1 ratio using the protocol described below (this is the protocol we followed on Wednesday in which everything is conducted under nitrogen in a Schlenk line)
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#Add deionized H<sub>2</sub>O to the round bottom flask until the solution is 5mL. Seal the round bottom flask with a septum.
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-A 5mL solution of 30mM FeSO4·7H2O, 100 mM KOH, and 400 mM KNO3 will be prepared with hemoglobin. The mixture will be incubated at room temperature for 5min while bubbling nitrogen gas and placed in a round bottom with a septum. The reaction mixture will be heated using a hot water bath to 90 degrees celcius for 4 hours.  
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#Using a Schlenk line, de-gas the solution using nitrogen. De-gas the solution using nitrogen gas at room temperature for 5 minutes.
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#Heat the reaction mixture to 90°C using a hot water bath for 4 hours. During this time, the reaction mixture should have a continuous flow of nitrogen gas.
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<br>
*Note: A previously prepared stock solution of 15mM BSA was used for the preparation.  
*Note: A previously prepared stock solution of 15mM BSA was used for the preparation.  
Table 1. Preparation of BSA:MagnetiteNP solution  
Table 1. Preparation of BSA:MagnetiteNP solution  

Revision as of 20:41, 3 February 2014

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Objectives

  1. Prepare a BSA-Magnetite solution with the purpose of synthesizing BSA fibers with magnetite nanoparticles incorporated in the fiber.
  2. Analyze protein-AuNP solutions prepared on 01/28/2014 using UV/vis.
  3. Analyze protein-AuNP-Magnetite solutions prepared on 01/28/2014 using UV/vis.


Preparation of BSA-Magnetite solution

  • The following protocol was adapted from the following reference[[1]]

General Protocol:

  1. Prepare a stock solution of BSA in a volumetric flask using deionized H2O.
  2. Prepare a 0.3M stock solution of FeSO4*7H2O in a 10mL volumetric flask using deionized H2O.
  3. Prepare a 1M stock solution of KOH in a 10mL volumetric flask using deionized H2O.
  4. Prepare a 0.8M stock solution of KNO3 in a 50mL volumetric flask using deionized H2O.
  5. Add the necessary amount of BSA for the desired ratio of BSA to Magnetite to a 10mL round bottom flask.
  6. Add the appropriate volume of FeSO4*7H2O to each test tube so that the solution has a concentration of 0.03M FeSO4*7H2O.
  7. Add the appropriate volume of KOH to each test tube so that the solution has a concentration of 0.1M KOH.
  8. Add the appropriate volume of KNO3 to each test tube so that the solution has a concentration of 0.4M KNO3.
  9. Add deionized H2O to the round bottom flask until the solution is 5mL. Seal the round bottom flask with a septum.
  10. Using a Schlenk line, de-gas the solution using nitrogen. De-gas the solution using nitrogen gas at room temperature for 5 minutes.
  11. Heat the reaction mixture to 90°C using a hot water bath for 4 hours. During this time, the reaction mixture should have a continuous flow of nitrogen gas.


  • Note: A previously prepared stock solution of 15mM BSA was used for the preparation.

Table 1. Preparation of BSA:MagnetiteNP solution

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)
1300.0001153850.7354058980.50.5004459512.50.764148151

UV/vis

General Protocol:

  1. Place 1mL of each ratio in an 1.5mL eppendorf tube.
  2. Centrifuge at maximum speed for 30 minutes.
  3. Take a UV-Vis spectrum of each sample from 200nm to 800nm.
  4. After taking the spectrum, store the sample in a fresh eppy tube in an upright position.


Figure 1. Corrected Absorbance Spectra of BSA-AuNP solutions (...ratios)

Notes


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