|
|
(6 intermediate revisions by one other user not shown) |
Line 2: |
Line 2: |
| |- | | |- |
| |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> |
| |style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html> </html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}} | | |style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} |
| |- | | |- |
| | 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]]
| |
|
| |
|
|
| |
|