User:Catherine Koenigsknecht/Notebook/Experimental Biological Chemistry/2013/02/06: Difference between revisions

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|style="background-color: #EEE"|[[Image:BDLlogo_notext_lr.png|128px]]<span style="font-size:22px;"> Biomaterials Design Lab</span>
|style="background-color: #EEE"|[[Image:BDLlogo_notext_lr.png|128px]]<span style="font-size:22px;"> Biomaterials Design Lab</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>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
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Learn how to maintain an OpenWetWare Notebook.
Learn how to maintain an OpenWetWare Notebook.


==Description==
#UVProbe was opened.
##Window > 1. Kinetics
##Methods Icon
###Wavelength: 265nm
###Duration: 300 seconds (5minutes)
###OK
#Shimadzu CPS-Controller was set to 25°C.
##wait for the temperature to raise to 25°C
#place the sample in the cell and click start.


*Phosphate buffer was used as a blank and was used to create the baseline.
*Kinetics was run at 265nm using 2.38mL phosphate buffer, 600μL of 500μM stock adenosine, and 20μL ADA (as outlined in the table below).
**The velocity was determined by clicking Operations > Activity Table > dAbs. The average of the dAbs values is the average slope (velocity).




*5mM adenosine stock solution was too concentrated and exceeded the capacity of the UV-Vis therefore it was diluted:
**150 uM adenosine stock solution was used; (stock) 150uL 5mM stock + 850 uL buffer =750 uM ->use .6mL new stock for ADA assay
**200 uM adenosine stock solution was used: (stock) 200 uL 5mM stock+800 uL buffer = 1000uM ->use 0.6mL new stock for ADA assay
**This table is the volumes of 5mM adenosine and phosphate buffer used to make the new adenosine stock solutions.
***[[Image:Screen_Shot_2013-02-05_at_9.01.20_PM.png]]
**This table is the updated table of the volumes and concentrations of solutions that will be added in the cuvette.
***[[Image:Screen_Shot_2013-02-05_at_9.01.33_PM.png]]
==Data==
{| {{table}}
| align="center" style="background:#f0f0f0;"|'''Concentration of Adenosine (uM)'''
| align="center" style="background:#f0f0f0;"|'''Average Velocity over 30s (nm/sec)'''
|-
| 200||0.00021
|-
| 100||0.000183333
|-
| 40||0.00012
|-
| 16||0.0001
|-
| 6.4||0.000047
|-
| 2.56||0.000033
|}
{| {{table}}
| align="center" style="background:#f0f0f0;"|'''1/[adenosine] (1/uM)'''
| align="center" style="background:#f0f0f0;"|'''1/[velocity] (sec/nm)'''
|-
| 0.005||4761.904762
|-
| 0.01||5454.545455
|-
| 0.025||8333.333333
|-
| 0.0625||10000
|-
| 0.15625||21276.59574
|-
| 0.390625||30303.0303
|}
*Lineweaver Burke Plot:
[[Image:ConcentrationvsVelocity-1.png]]
*Concentration Adenosine vs Velocity:
[[Image:LineweaverBurkePlot-1.png]]
{| {{table}}
| -1/Vmax||6163.1
|-
| '''Vmax'''||'''0.000162256 nm/sec'''
|-
| -1/Km||-0.093
|-
| '''Km'''||'''10.75 uM'''
|}
==Notes==
This area is for any observations or conclusions that you would like to note.




Latest revision as of 22:25, 26 September 2017

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