User:Zachary I. Mendel/Notebook/Zacks Notebook/2014/02/11

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==Notes==
==Notes==
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*the nanoparticles with HRP were functional
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*In the 10^-11 dillution, we observed a color change from red initially to a much lighter solution
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*In our calculations we decided to use teh 10^-11 and water as our blank, so we could subtract protein + water.
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<br>
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Protein, OBD, H202
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*For each sample run in UV-Vis  there was 0.225ml HRP, 2.25ml OBD stock and 0.75ml hydrogen peroxide stock
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<br>
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Revision as of 16:35, 11 February 2014

Project name Main project page
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February 11th

Goals

  • We have to redo the dopamine addition that we did last Wednesday
  • We are going to test the activity of citrate-HRP-AuNPs using O2 binding assays


Procedures



Activity of citrate-HRP-AuNPs

  • We are going to use an assay provided by Dr. Hartings
    • The assay uses a reagent that changes color when HRP is added in addition to hydrogen peroxide
    • The hydrogen peroxide alone will change the color of the solution, so we will be comparing hydrogen peroxide alone with hydrogen peroxide and AuNPs
  • We also are going to run UV Vis of the samples to see how the absorption spectra changes over time


Dopamine Coating

  1. Dissolve dopamine in 100mL Tris Buffer to have a 10mM solution at pH=8.5
  2. Mix 17mL of AuNP solution with 17mL of the dopamine solution
  3. Stir under air at room temperature for 1 hour
  4. Purify by centrifuging and then redisperse in water.
  5. Filtrate through syringe filters with cellulose acetate membranes with pore size of 1um
  6. Make 2 solutions.
    1. to be stored over night on lab bench
    2. to be stored over night in the refrigerator

Notes

  • the nanoparticles with HRP were functional
  • In the 10^-11 dillution, we observed a color change from red initially to a much lighter solution
  • In our calculations we decided to use teh 10^-11 and water as our blank, so we could subtract protein + water.


Protein, OBD, H202

  • For each sample run in UV-Vis there was 0.225ml HRP, 2.25ml OBD stock and 0.75ml hydrogen peroxide stock



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