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Experimental Biological Chemistry: Fall 2013 <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
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October 16, 2013

Objective

  • Test the activity of HRP-AuNPs today for the catalytic conversion of luminol

Description

Reactions (Amounts in Cuvette)

  • Luminol: final concentration so A(330)<1
    • Stock: 1.46mM
    • Make 20x dilution
    • Final: 0.073mM
  • H2O2: final concentration 50x [Luminol]
    • Stock: 44.29mM
    • 50×0.073mM=3.65
    • Final: 3.65mM
  • HRP-NPs: use 100uL of 60:1 solution
    • [Au]=0.25mM
    • [HRP]=4.167uM

<br.>

Kinetics

  • Run with H2O2, luminol, buffer solution, adding 100uL HRP-NPs to start reaction

<br.>

Remake BSA-AuNPs

  1. Calculations:
    • Stock [HAuCl4]: 2.33mM
    • Stock [BSA]:15.0uM
    • Final [HAuCl4]= 90×[BSA]=90×15.0uM=1350μM=1.350mM=final [HAuCl4]
      • (2.33mM HAuCl4 initial)(1mL solution)=(1.350mM HAuCl4 final)(V)
      • V=1.73mL BSA<br.>
    • Combine 1mL HAuCl4 and 1.73mL BSA, dilute with distilled water up to 10mL
  2. Transfer solution to a test tube and cap with aluminum foil
  3. Heat in oven at 80C
  4. Transfer solution to a plastic falcon tube

<br.>

Reactions: with 1mL HRP-NPs

  • Same concentrations, 1mL less buffer for 1mL HRP-NPs

Data

  • Very slow kinetics observed for addition of 100uL HRP-NPs

Notes


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