User:Javier Vinals Camallonga/Notebook/Javier Vinals notebook/2014/04/02: 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.  
* Run UV-Vis and AA on room temperature 30:1 lysozyme-AuNP samples from 3/26/14.
* Run conductivity on pure variables at room temperature.
 
 
==Procedure==
 
===Conductivity Measurement of Pure Variables at Room Temperature===
[[Image:4.2.cod.jac.png|800px|]]
 
 
 
===Atomic Absorption Preparation===
 
'''Creating the Gold Stock Solutions'''
# Add 50 μL of HAuCl<sub>4</sub>·3H<sub>2</sub>O and 4950 μL of distilled water to a Falcon tube, for a final concentration of 10 μg/mL Au.
# Add 100 μL of HAuCl<sub>4</sub>·3H<sub>2</sub>O and 4900 μL of distilled water to a Falcon tube, for a final concentration of 20 μg/mL Au.
# Add 150 μL of HAuCl<sub>4</sub>·3H<sub>2</sub>O and 4850 μL of distilled water to a Falcon tube, for a final concentration of 30 μg/mL Au.
# Add 200 μL of HAuCl<sub>4</sub>·3H<sub>2</sub>O and 4800 μL of distilled water to a Falcon tube, for a final concentration of 40 μg/mL Au.
# Add 250 μL of HAuCl<sub>4</sub>·3H<sub>2</sub>O and 4750 μL of distilled water to a Falcon tube, for a final concentration of 50 μg/mL Au.
 
 
 
'''Atomic Absorption Samples'''
 
Solutions with the following Au:lysozyme ratio at room temperature were run on the AA:
* 30:1 lysozyme-AuNP with 0.03 M MgCl<sub>2</sub>, CaCl<sub>2</sub>, NaCl, KCl, MES, citric acid (0.0002316 M 2,2 bipyridine)
* 30:1 lysozyme-AuNP with 0.06 M MgCl<sub>2</sub>, CaCl<sub>2</sub>, NaCl, KCl, MES, citric acid (0.0004544 M 2,2 bipyridine)
* 30:1 lysozyme-AuNP with 0.09 M MgCl<sub>2</sub>, CaCl<sub>2</sub>, NaCl, KCl, MES, citric acid (0.0006772 M 2,2 bipyridine)
 
==Figures==
[[Image:4.2.14.aa.jac.png|850px|]]


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.




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Entry title

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Objective

  • Run UV-Vis and AA on room temperature 30:1 lysozyme-AuNP samples from 3/26/14.
  • Run conductivity on pure variables at room temperature.


Procedure

Conductivity Measurement of Pure Variables at Room Temperature


Atomic Absorption Preparation

Creating the Gold Stock Solutions

  1. Add 50 μL of HAuCl4·3H2O and 4950 μL of distilled water to a Falcon tube, for a final concentration of 10 μg/mL Au.
  2. Add 100 μL of HAuCl4·3H2O and 4900 μL of distilled water to a Falcon tube, for a final concentration of 20 μg/mL Au.
  3. Add 150 μL of HAuCl4·3H2O and 4850 μL of distilled water to a Falcon tube, for a final concentration of 30 μg/mL Au.
  4. Add 200 μL of HAuCl4·3H2O and 4800 μL of distilled water to a Falcon tube, for a final concentration of 40 μg/mL Au.
  5. Add 250 μL of HAuCl4·3H2O and 4750 μL of distilled water to a Falcon tube, for a final concentration of 50 μg/mL Au.


Atomic Absorption Samples

Solutions with the following Au:lysozyme ratio at room temperature were run on the AA:

  • 30:1 lysozyme-AuNP with 0.03 M MgCl2, CaCl2, NaCl, KCl, MES, citric acid (0.0002316 M 2,2 bipyridine)
  • 30:1 lysozyme-AuNP with 0.06 M MgCl2, CaCl2, NaCl, KCl, MES, citric acid (0.0004544 M 2,2 bipyridine)
  • 30:1 lysozyme-AuNP with 0.09 M MgCl2, CaCl2, NaCl, KCl, MES, citric acid (0.0006772 M 2,2 bipyridine)

Figures



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