User:Megan L. Channell/Notebook/Horseradish/2013/10/08

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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
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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: #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>}}
|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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==Entry title==
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* Insert content here...
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==Objective==
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Measure and observe ADA kinetics without inhibitor.  This work will be the basis of our comparison to ADA-AuNPtunrover studies.
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From [[User:Matt_Hartings/Notebook/AU_Biomaterials_Design_Lab/2013/10/08 | Dr. Hartings]].
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==Procedure==
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# Make a 40μM solution of adenosine in buffer (50mM phosphate buffer, pH 7.4)
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# Go to Dr. Hartings lab for enzyme kinetics measurements.
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## Add 3mL of adenosine solution to the cuvette
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## Start your kinetics measurement
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### 1ms integration (on front panel)
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### 10 scan average (on front panel)
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### Set "Save the first available scan every" to 15 seconds (after clicking File>Save)
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### Set "Stop after this amount of time" to 10 minutes (after clicking File>Save)
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### Set "File Type" to Tab Delimited
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### Give the files a directory and a name
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### Click accept
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### Just before 1 minute add 30ul of 0.01u/mL ADA
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*taken from [[User:Matt_Hartings/Notebook/AU_Biomaterials_Design_Lab/2013/10/08 | Dr. Hartings]]
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#How we made the 40μM solution of adenosine in buffer
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##.1063 grams of adenosine were dissolved in a 10mL volumetric flask. Molarity: ~40mM
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## the solution was mixed to ensure homogenity
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##9.98 μL of 40mM solution were diluted up to 10mL. Molarity: 40μM
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==Data==
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[[Image:Cmj_10.09.13_ADAnoinhibitor.png]]
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==Notes==
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[[Category:CHEM571]]
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[[Category:Adenosine]]
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[[Category:Inosine]]
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Revision as of 15:33, 10 October 2013

Biomaterials Design Lab Main project page
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Objective

Measure and observe ADA kinetics without inhibitor. This work will be the basis of our comparison to ADA-AuNPtunrover studies. From Dr. Hartings.


Procedure

  1. Make a 40μM solution of adenosine in buffer (50mM phosphate buffer, pH 7.4)
  2. Go to Dr. Hartings lab for enzyme kinetics measurements.
    1. Add 3mL of adenosine solution to the cuvette
    2. Start your kinetics measurement
      1. 1ms integration (on front panel)
      2. 10 scan average (on front panel)
      3. Set "Save the first available scan every" to 15 seconds (after clicking File>Save)
      4. Set "Stop after this amount of time" to 10 minutes (after clicking File>Save)
      5. Set "File Type" to Tab Delimited
      6. Give the files a directory and a name
      7. Click accept
      8. Just before 1 minute add 30ul of 0.01u/mL ADA
  1. How we made the 40μM solution of adenosine in buffer
    1. .1063 grams of adenosine were dissolved in a 10mL volumetric flask. Molarity: ~40mM
    2. the solution was mixed to ensure homogenity
    3. 9.98 μL of 40mM solution were diluted up to 10mL. Molarity: 40μM

Data

Image:Cmj_10.09.13_ADAnoinhibitor.png

Notes




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