User:Javier Vinals Camallonga/Notebook/Javier Vinals notebook/2013/10/15: Difference between revisions

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==Objective==
==Objective==
Today we'll be determining the molar absorptivities of two different molecules, [http://en.wikipedia.org/wiki/Adenosine adenosine] and [http://en.wikipedia.org/wiki/Inosine inosine]. The data that we generate today will be important when we study [http://en.wikipedia.org/wiki/Adenosine_deaminase adenosine deaminase] (ADA), which converts adenosine to inosine. The difference between these two molecules is that adenosine contains a primary amine whereas inosine contains a carboxy group. Overexpression of this protein causes anemia in humans. A shortage of this protein can lead to severe immuno-defficiency.  
Today we are going to be making HRP-AuNPs for enzymatic studies.


Adenosine and inosine have different absorption spectra. We will be observing changes in UV-Vis spectra to determine changes in concentration of both adenosine and inosine. In order to do this, we will need to know the molar absorptivity (ε) of both of these molecules. Just as each molecule has a characteristic absorption at each wavelength, this (per-wavelength) absorption can be quantified by a molar absorptivity. Or ... for a given concentration a molecule will absorb a very specific amount of light at a precise wavelength. A molecule doesn't have just one molar absorptivity; there is a molar absorptivity to describe each wavelength in a molecular absorbance spectrum.
==Description==
# Make 4 solutions of HRP-AuNPs
## Au:HRP ratio - 30:1 HRP-Au (final concentration of Au - 0.25mM; total volume - 10mL)
## Au:HRP ratio - 60:1 HRP-Au (final concentration of Au - 0.25mM; total volume - 10mL)
## Au:HRP ratio - 90:1 HRP-Au (final concentration of Au - 0.25mM; total volume - 10mL)
## Au:HRP ratio - 120:1 HRP-Au (final concentration of Au - 0.25mM; total volume - 10mL)
 
==Data==
Stock solutions
# Gold
## 25.0mg HAuCl<sub>4</sub>·H<sub>2</sub>O in 25.0mL water --> 2.94mM
# HRP
## 17.9mg HRP in 25.0mL water --> 16.27μM




Revision as of 11:40, 15 October 2013

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Objective

Today we are going to be making HRP-AuNPs for enzymatic studies.

Description

  1. Make 4 solutions of HRP-AuNPs
    1. Au:HRP ratio - 30:1 HRP-Au (final concentration of Au - 0.25mM; total volume - 10mL)
    2. Au:HRP ratio - 60:1 HRP-Au (final concentration of Au - 0.25mM; total volume - 10mL)
    3. Au:HRP ratio - 90:1 HRP-Au (final concentration of Au - 0.25mM; total volume - 10mL)
    4. Au:HRP ratio - 120:1 HRP-Au (final concentration of Au - 0.25mM; total volume - 10mL)

Data

Stock solutions

  1. Gold
    1. 25.0mg HAuCl4·H2O in 25.0mL water --> 2.94mM
  2. HRP
    1. 17.9mg HRP in 25.0mL water --> 16.27μM



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