Difference between revisions of "User:James C. Schwabacher/Notebook/CHEM-571/2013/10/16"

From OpenWetWare
Jump to: navigation, search
(fix raw html notebook nav)
(4 intermediate revisions by one other user not shown)
Line 2: Line 2:
|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>}}
|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]]&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}}
| colspan="2"|
| colspan="2"|
Line 42: Line 42:
[[Image:Cmj_HRPKineticsUV-Vis_10.16.2013.png ]]
*Note: Spectra overlap for a total elapsed time of 19 minutes.

Latest revision as of 22:29, 26 September 2017

BDLlogo notext lr.png Biomaterials Design Lab Report.pngMain project page
Resultset previous.pngPrevious entry      Next entryResultset next.png


To test the activity of our HRP-NPs for the catalytic conversion of luminol.


Creating the Luminol-H2O2 Solution

  1. Add 969.61 μL of the luminol stock and 1.598 mL of the hydrogen peroxide stock to a 10 mL volumetric flask.
  2. Dilute the solution to 10 mL with buffer.

Enzyme Kinetics Measurement

  1. Add 2 mL of the luminol-H2O2 solution to a cuvette.
  2. Start kinetics measurement:
  • 1 ms integration
  • 10 scan average
  • Set "Save the first available scan every" to 15 seconds
  • Set "Stop after this amount of time" to 10 minutes
  • Set "File Type" to Tab Delimited
  • Just before 1 minute, add 1 mL of 60:1 Au:HRP solution.

BSA-AuNP Synthesis

  1. Add 1.00 mL of 2.33 mM gold solution (HAuCL4) to a 10 mL volumetric flask.
  2. Add 1.73 mL BSA (90x more gold than BSA) to the volumetric flask.
  3. Dilute the solution up to 10 mL with deionized water.
  4. Transfer the solution to a test tube and cap with aluminum foil.
  5. Heat in the oven at 80°C for four hours.
  6. Transfer the solution to a plastic falcon tube.

Remaking the Solution of Gold and HRP

  1. For 60:1 Au:HRP solution, 2.56 mL HRP stock and 850.34 μL of gold stock were added to a 10 mL volumetric flask. The solution was diluted up to 10 mL with deionized water.
  2. The 10 mL solution of gold and HRP was placed into a test tube.
  3. The test tube was placed in the oven at 80°C for four hours.


Cmj HRPKineticsUV-Vis 10.16.2013.png

  • Note: Spectra overlap for a total elapsed time of 19 minutes.

Cmj HRPKinetics 520overtime 10.16.2013.png


  • The buffer was 5.1 mM Tris at pH 8.
  • The stock concentration of luminol was 1.46 mM.
  • The stock concentration of hydrogen peroxide was 44.29 mM.
  • The stock concentration of gold (HAuCL4)was 2.33 mM.
  • The stock concentration of BSA was 15 μM.
  • The stock solution of gold was 2.94 mM.
  • The stock solution of HRP was 16.27 μM.