User:Carly M. Montanero/Notebook/CHEM-571/2013/10/16: Difference between revisions

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:* Set "File Type" to Tab Delimited
:* Set "File Type" to Tab Delimited
:* Just before 1 minute, add 1 mL of 60:1 Au:HRP solution.
:* Just before 1 minute, add 1 mL of 60:1 Au:HRP solution.
'''BSA-AuNP Synthesis'''
# Add 1.00 ml of 2.54 mM gold solution (HAuCL<sub>4</sub>) to a 10 ml volumetric flask.
# Add 1.81 BSA (90x more gold than BSA) to the volumetric flask.
# Add 7.19 ml deionized water to the volumetric flask.
# Transfer the solution to a test tube and cap with aluminum foil.
# Heat in the oven at 80°C for three hours.
# Transfer the solution to a plastic falcon tube.


==Notes==
==Notes==

Revision as of 11:52, 16 October 2013

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Objective

To test the activity of our HRP-NPs for the catalytic conversion of luminol.

Procedure

Creating the Luminol-H2O2 Solution

  1. Add 969.61 μL of the luminol stock and 1.598 mL of the hydrogen peroxide stock to a 10 mL volumetric flask.
  2. Dilute the solution to 10 mL with buffer.

Enzyme Kinetics Measurement

  1. Add 2 mL of the luminol-H2O2 solution to a cuvette.
  2. Start kinetics measurement:
  • 1 ms integration
  • 10 scan average
  • Set "Save the first available scan every" to 15 seconds
  • Set "Stop after this amount of time" to 10 minutes
  • Set "File Type" to Tab Delimited
  • Just before 1 minute, add 1 mL of 60:1 Au:HRP solution.

BSA-AuNP Synthesis

  1. Add 1.00 ml of 2.54 mM gold solution (HAuCL4) to a 10 ml volumetric flask.
  2. Add 1.81 BSA (90x more gold than BSA) to the volumetric flask.
  3. Add 7.19 ml deionized water to the volumetric flask.
  4. Transfer the solution to a test tube and cap with aluminum foil.
  5. Heat in the oven at 80°C for three hours.
  6. Transfer the solution to a plastic falcon tube.

Notes

  • The buffer was 5.1 mM Tris at pH 8.
  • The stock concentration of luminol was 1.46 mM.
  • The stock concentration of hydrogen peroxide was 44.29 mM.


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