User:Michael F. Nagle/Notebook/Chem 571/2013/02/12: Difference between revisions

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==Objective==
 
Repeat ADA Assay 2 times for additional data


==Description==
==Description==

Revision as of 14:25, 8 May 2013

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Description

  • Using procedure for UV-Vis from 2/6/13 two assays were performed using Adenosine and ADA in phosphate buffer
  • 5mM adenosine stock solution was too concentrated and exceeded the capacity of the UV-Vis therefore it was diluted:
    • Dilution Tables are located below

Concentration Tables

This table is the volumes of 5mM adenosine and phosphate buffer used to make the new adenosine stock solutions.

[Old Adenosine Stock Solution] (uM) Volume Adenosine Stock Solution added (uL) Volume Adenosine Phosphate Buffer added (uL) [New Adenosine Stock Solution] (uL) Volume New Adenosine Stock Solution (mL)
5000 200 800 1000 1
5000 150 850 750 1
5000 100 900 500 1
5000 40 960 200 1
5000 16 984 80 1
5000 6.4 993.6 32 1

This table is the updated table of the volumes and concentrations of solutions that will be added in the cuvette.

[ADA Stock Solution] (U/mL) Volume ADA Stock Solution Added (mL) Final [ADA] (U/mL) [Adenosine Stock Solution] (mL) Volume Adenosine Stock Solution Added (mL) Final [Adenosine] (uM) Volume 0.05 M Phosphate Buffer Added (mL) Total Volume (mL)
0.5 0.02 0.00333 1000 0.6 200 2.38 3
0.5 0.02 0.00333 750 0.6 150 2.38 3
0.5 0.02 0.00333 500 0.6 100 2.38 3
0.5 0.02 0.00333 200 0.6 40 2.38 3
0.5 0.02 0.00333 80 0.6 16 2.38 3
0.5 0.02 0.00333 32 0.6 6.4 2.38 3

Data

Run#1

Concentration of Adenosine (uM) Average Velocity over 180s (nm/sec)
200 0.000116022
150 0.000192818
100 0.000139227
40 5.55556E-06
16 6.68508E-05
6.4 0.000039
1/Concentration of Adenosine (uM) 1/Velocity (nm/sec)
0.005 8619.047619
0.006666667 5186.246418
0.01 7182.539683
0.025 180000
0.0625 14958.67769
0.15625 25641.02564


Run#2

Concentration of Adenosine (uM) Average Velocity over 180s (nm/sec)
200 0.00021989
150 0.000147514
100 0.000144751
40 0.00010884
16 8.12155E-05
6.4 3.8674E-05
1/Concentration of Adenosine (uM) 1/Velocity (nm/sec)
0.005 4547.738693
0.006666667 6779.026217
0.01 6908.396947
0.025 9187.817259
0.0625 12312.92517
0.15625 25857.14286



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