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
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Kinetics
- Run with H2O2, luminol, buffer solution, adding 100uL HRP-NPs to start reaction
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Remake BSA-AuNPs
- 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
- Transfer solution to a test tube and cap with aluminum foil
- Heat in oven at 80C
- Transfer solution to a plastic falcon tube
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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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