User:Moira M. Esson/Notebook/CHEM-571/2013/10/08: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
(Autocreate 2013/10/08 Entry for User:Moira_M._Esson/Notebook/CHEM-571)
 
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. ##### -->
==Entry title==
==Objectives==
* Insert content here...
#Monitor the turnover kinetics of adenosine to inosine catalyzed by adenosine deaminase (ADA)
<br>
==Kinetics==
'''General Protocol''':
#Prepare a 40μM solution of adenosine in 50mM phosphate pH 7.4 buffer.
#*Note: This solution was prepared via dilution of a 0.0123M ADA stock solution
#Add 3mL of 40μM adenosine to a cuvette with a stir bar
#*Note: All analysis should be carrier out in a temperature controlled environment. The temperature was kept at 25°C
#Allow the solution to reach a constant 25°C
#Begin taking absorbance spectra (take a spectrum every 15seconds)
#After 1min, add 30μL 1.1units/mL ADA
#Allow the reaction to run for 10 minutes, taking a spectrum every 15seconds.
<br>
'''Sample Preparation''':
Table 1. Preparation of 0.0123M stock solution
<br>
{| {{table}}
| align="center" style="background:#f0f0f0;"|Mass of adenosine (g)
| align="center" style="background:#f0f0f0;"|0.0165
|-
| Volume of buffer(mL)||5
|}
<br>
Table 2. Preparation of 40μM adenosine solution
<br>
{| {{table}}
| align="center" style="background:#f0f0f0;"|Volume of stock(mL)
| align="center" style="background:#f0f0f0;"|0.0098
|-
| Volume of buffer(mL)||2.9902
|}




Revision as of 10:22, 8 October 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>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

Objectives

  1. Monitor the turnover kinetics of adenosine to inosine catalyzed by adenosine deaminase (ADA)


Kinetics

General Protocol:

  1. Prepare a 40μM solution of adenosine in 50mM phosphate pH 7.4 buffer.
    • Note: This solution was prepared via dilution of a 0.0123M ADA stock solution
  2. Add 3mL of 40μM adenosine to a cuvette with a stir bar
    • Note: All analysis should be carrier out in a temperature controlled environment. The temperature was kept at 25°C
  3. Allow the solution to reach a constant 25°C
  4. Begin taking absorbance spectra (take a spectrum every 15seconds)
  5. After 1min, add 30μL 1.1units/mL ADA
  6. Allow the reaction to run for 10 minutes, taking a spectrum every 15seconds.


Sample Preparation: Table 1. Preparation of 0.0123M stock solution

Mass of adenosine (g) 0.0165
Volume of buffer(mL) 5


Table 2. Preparation of 40μM adenosine solution

Volume of stock(mL) 0.0098
Volume of buffer(mL) 2.9902



Retrieved from "https://openwetware.org/mediawiki/index.php?title=User:Moira_M._Esson/Notebook/CHEM-571/2013/10/08&oldid=736019"