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| |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}}}}]]}} |
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| ==Procedure==
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| #UVProbe was opened.
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| ##Window > 1. Kinetics
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| ##Methods Icon
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| ###Wavelength: 265nm
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| ###Duration: 300 seconds (5minutes)
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| ###OK
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| #Shimadzu CPS-Controller was set to 25°C.
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| ##wait for the temperature to raise to 25°C
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| #place the sample in the cell and click start.
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| *Phosphate buffer was used as a blank and was used to create the baseline.
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| *Kinetics was run at 265nm using 2.38mL phosphate buffer, 600μL of 500μM stock adenosine, and 20μL ADA (as outlined in the table below).
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| **The velocity was determined by clicking Operations > Activity Table > dAbs. The average of the dAbs values is the average slope (velocity).
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| *5mM adenosine stock solution was too concentrated and exceeded the capacity of the UV-Vis therefore it was diluted:
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| **150 uM adenosine stock solution was used; (stock) 150uL 5mM stock + 850 uL buffer =750 uM ->use .6mL new stock for ADA assay
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| **200 uM adenosine stock solution was used: (stock) 200 uL 5mM stock+800 uL buffer = 1000uM ->use 0.6mL new stock for ADA assay
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| **This table is the volumes of 5mM adenosine and phosphate buffer used to make the new adenosine stock solutions.
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| ***[[Image:Screen_Shot_2013-02-05_at_9.01.20_PM.png]]
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| **This table is the updated table of the volumes and concentrations of solutions that will be added in the cuvette.
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| ***[[Image:Screen_Shot_2013-02-05_at_9.01.33_PM.png]]
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| ==Data==
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| {| {{table}}
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| | align="center" style="background:#f0f0f0;"|'''Concentration of Adenosine (uM)'''
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| | align="center" style="background:#f0f0f0;"|'''Average Velocity over 30s (nm/sec)'''
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| | 200||0.00021
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| | 100||0.000183333
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| |-
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| | 40||0.00012
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| | 16||0.0001
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| |-
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| | 6.4||0.000047
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| |-
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| | 2.56||0.000033
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| |}
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| {| {{table}}
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| | align="center" style="background:#f0f0f0;"|'''1/[adenosine] (1/uM)'''
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| | align="center" style="background:#f0f0f0;"|'''1/[velocity] (sec/nm)'''
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| | 0.005||4761.904762
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| | 0.01||5454.545455
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| |-
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| | 0.025||8333.333333
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| |-
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| | 0.0625||10000
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| |-
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| | 0.15625||21276.59574
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| |-
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| | 0.390625||30303.0303
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| |}
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| *[[Image:Graphsmn1.jpg]]
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| {| {{table}}
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| | -1/Vmax||6163.1
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| | '''Vmax'''||'''0.000162256 nm/sec'''
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| | -1/Km||-0.093
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| | '''Km'''||'''10.75 uM'''
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| |}
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