User:Mary Mendoza/Notebook/CHEM572 Exp. Biological Chemistry II/2013/02/12: Difference between revisions
From OpenWetWare
Mary Mendoza (talk | contribs) (Autocreate 2013/02/12 Entry for User:Mary_Mendoza/Notebook/CHEM572_Exp._Biological_Chemistry_II) |
Mary Mendoza (talk | contribs) |
||
Line 6: | Line 6: | ||
| colspan="2"| | | colspan="2"| | ||
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### --> | <!-- ##### DO NOT edit above this line unless you know what you are doing. ##### --> | ||
== | ==ADA Kinetic Assay== | ||
* | *The assay runs were continued from last week's, [[User:Mary Mendoza/Notebook/CHEM572 Exp. Biological Chemistry II/2013/02/05|02/05/2013]] calculated values. | ||
*The objective of performing the assay is to obtain the rate of the enzymatic reaction of adenosine into inosine by ADA. | |||
*Baseline 200-400nm using pH 7.4, 0.05M phosphate buffer | |||
*Temperature control: 25°C | |||
*Phosphate buffer in reference cell | |||
*Ran kinetics on 250uM solution (20uL ADA, 600uL 1250uM Adenosine, and 2.38mL pH7.4, 0.005M phosphate buffer) for 5 minutes at 265nm. | |||
**absorbance exceeded 4.00, so the assay was stopped and a new solution was prepared. | |||
*Ran kinetics on 2.56uM solution (20uL ADA, 600uL 12.8uM Adenosine, and 2.38mL pH7.4, 0.005M phosphate buffer) for 5 minutes at 265nm. | |||
**dAbs was obtained and plotted vs. time in excel | |||
*Ran kinetics on 6.4uM solution (20uL ADA, 600uL 32uM Adenosine, and 2.38mL pH7.4, 0.005M phosphate buffer) for 5 minutes at 265nm. | |||
**dAbs was obtained and plotted vs. time in excel | |||
*Ran kinetics on 16uM solution (20uL ADA, 600uL 80uM Adenosine, and 2.38mL pH7.4, 0.005M phosphate buffer) for 5 minutes at 265nm. | |||
**dAbs was obtained and plotted vs. time in excel | |||
*The following table describes the volumes and concentrations of the components added to the cuvette. | |||
[[Image:Data_Table.JPG|center]] | |||
Revision as of 22:08, 12 February 2013
Project name | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> |
ADA Kinetic Assay
|