User:Michael F. Nagle/Notebook/Chem 571/2012/09/25: Difference between revisions

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==Data==
==Data==
{| border="1"
|+ Amount of Tris stock and water in each tube
! scope="col" |'''Tris Buffer concentration'''
! scope="col" |'''amount of water (mL)'''
! scope="col" |'''amount of tris (mL)'''
|-
| .05 mM||0.9938||0.0625
|-
| .5 mM||0.9875||0.0125
|-
| 5 mM||0.975||0.025
|-
| 50 mM||0.95||0.05
|-
| 100 mM||0.9||0.1
|-
| 200 mM||0.8||0.2
|-
| 500 mM||0.5||0.5
|-
| 1 M||0||1




Revision as of 12:28, 1 October 2012

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  1. Protein extraction
    1. Cells were removed from freezer and warmed to room temperature in a water bath.
    2. Cells were sonicated
      1. Each vial was placed in an ice bath for 30 seconds
      2. Sonication device was placed in each vial for 30 seconds
      3. This was repeated 3 times for each vial.
    3. Cells were centrifuged at 18000rpm and 4°C for 2 hours to seperate ADA from E. Coli
    4. Binding and elution buffers were purified via vacuum filtration and a 450nm membrane filter.
    5. Vacuum filtration was used to seperate proteins from cells. ADA was collected in falcon vials and refridgerated.
  1. Solutions of Au/BSA at a mole ratio of 70 (at which fibers are not produced) with Tris at molarities of .05mM, .5mM, 5mM, 50mM, 100mM, 200mM, 500mM, and 1M went through two rounds each of UV/Vis.

Data

Amount of Tris stock and water in each tube
Tris Buffer concentration amount of water (mL) amount of tris (mL)
.05 mM 0.9938 0.0625
.5 mM 0.9875 0.0125
5 mM 0.975 0.025
50 mM 0.95 0.05
100 mM 0.9 0.1
200 mM 0.8 0.2
500 mM 0.5 0.5
1 M 0 1



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