User:Zachary I. Mendel/Notebook/Zacks Notebook/2013/10/16: Difference between revisions
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== | ==Objective== | ||
We are going to test the activity of our HRP-NPs today for the catalytic conversion of luminol | |||
==Description== | |||
# Add experimental record here. Include what, how, and why... | |||
==Data== | |||
<u>Stock Solution</u> | |||
# Buffer | |||
## 0.6140g Tris in 1L, pH set to 8 with HCl ---> 5.1mM | |||
# Luminol | |||
## Dissolve 12.9mg luminol in 300uL of DMSO | |||
## Add to 50mL buffer ---> 1.46mM | |||
# H2O2 | |||
## 312uL 30% H2O2 into 100mL buffer ---> Should be 45mM | |||
## Check absorption at 250 [http://www.bio.net/mm/methods/1995-January/023533.html source] (ε(250) = 16.69 M<sup>-1</sup>cm<sup>-1</sup>). A = 0.7392. [H2O2] = 44.29mM | |||
==Notes== | |||
Figure 1: Absorbance spectra of the catalytic activity of HRP-AuNPs | |||
<br> | |||
[[Image:AbsorbancespectracatalyticactivityHRPAuNPs.png]] | |||
- We measured the absorbance of the 1/20 luminol to see if it still absorbed. We found the absorbance peak to be less than 1.00, specifically around 0.300 | |||
Figure 2: Graph showing the absorbance spectra for citrate AuNP and HRP AuNP | |||
[[Image:HRP and Citrate AuNP.png]] | |||
Making the BSA AuNP solution | |||
-Add 1mL of the 2.54mM gold (HAuCl4) solution to a 10mL volumetric flask | |||
-Add an appropriate amount of BSA solution so that the final concentration of gold is 90X that of BSA | |||
- Calculations = ((0.00254 moles/L Gold)* (1L/1000ml)* (1ml)) / 90 = moles BSA = 2.82*10^-8 moles BSA | |||
[BSA] = 15.6μM | |||
- (2.82*10^-8 moles) * (1L/15.6*10^-6 moles) = 0.00180L or 1.8mL BSA to add to solution | |||
Add deionized water up to 10mL | |||
Transfer solution to a test tube and cap with aluminum foil | |||
Heat in oven at 80C for 3 hours | |||
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Revision as of 06:53, 28 October 2013
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ObjectiveWe are going to test the activity of our HRP-NPs today for the catalytic conversion of luminol Description
DataStock Solution
NotesFigure 1: Absorbance spectra of the catalytic activity of HRP-AuNPs
- We measured the absorbance of the 1/20 luminol to see if it still absorbed. We found the absorbance peak to be less than 1.00, specifically around 0.300 Figure 2: Graph showing the absorbance spectra for citrate AuNP and HRP AuNP Making the BSA AuNP solution -Add 1mL of the 2.54mM gold (HAuCl4) solution to a 10mL volumetric flask -Add an appropriate amount of BSA solution so that the final concentration of gold is 90X that of BSA - Calculations = ((0.00254 moles/L Gold)* (1L/1000ml)* (1ml)) / 90 = moles BSA = 2.82*10^-8 moles BSA [BSA] = 15.6μM - (2.82*10^-8 moles) * (1L/15.6*10^-6 moles) = 0.00180L or 1.8mL BSA to add to solution Add deionized water up to 10mL Transfer solution to a test tube and cap with aluminum foil Heat in oven at 80C for 3 hours |