User:Madeleine Y. Bee/Notebook/CHEM-571 2013F/2013/10/16: Difference between revisions

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**[Au]=0.25mM
**[Au]=0.25mM
**[HRP]=4.167uM
**[HRP]=4.167uM
<br.>
<u>Kinetics</u>
*Run with H<sub>2</sub>O<sub>2</sub>, luminol, buffer solution, adding 100uL HRP-NPs to start reaction
<br.>
<u>Remake BSA-AuNPs</u>
*See [http://openwetware.org/wiki/User:Matt_Hartings/Notebook/AU_Biomaterials_Design_Lab/2013/08/28 08/28/13] for procedure
#Calculations:
#*Stock [HAuCl<sub>4</sub>]: 2.33mM
#*Stock [BSA]:15.0uM
#*Final [HAuCl<sub>4</sub>]= 90×[BSA]=90×15.0uM=1350μM=1.350mM=final [HAuCl<sub>4</sub>]
#**(2.33mM HAuCl<sub>4</sub> initial)(1mL solution)=(1.350mM HAuCl<sub>4</sub> final)(V)
#**V=1.73mL BSA<br.>
#*Combine 1mL HAuCl<sub>4</sub> and 1.73mL BSA, dilute with distilled water up to 10mL
#Transfer solution to a test tube and cap with aluminum foil
#Heat in oven at 80C
#Transfer solution to a plastic falcon tube
<br.>
<u>Reactions: with 1mL HRP-NPs</u>
*Same concentrations, 1mL less buffer for 1mL HRP-NPs


===Data===
===Data===
*Very slow kinetics observed for addition of 100uL HRP-NPs


===Notes===
===Notes===

Revision as of 12:20, 16 October 2013

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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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