Difference between revisions of "User:Dhea Patel/Notebook/Experimental Biological Chemistry Notebook/2012/10/24"

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Please refer to table for volumes and concentrations of HRP, Luminol, Phenol, Buffer, and hydrogen peroxide used.
 
Please refer to table for volumes and concentrations of HRP, Luminol, Phenol, Buffer, and hydrogen peroxide used.
  
[[Image:20121024.png]]
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{| {{table}}
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| align="center" style="background:#f0f0f0;"|''''''
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| align="center" style="background:#f0f0f0;"|'''Volume and Stock Concentration Added to Cuvette'''
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| align="center" style="background:#f0f0f0;"|''''''
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| align="center" style="background:#f0f0f0;"|''''''
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| align="center" style="background:#f0f0f0;"|''''''
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| align="center" style="background:#f0f0f0;"|'''Volume Added to Cuvette [µL]'''
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| align="center" style="background:#f0f0f0;"|''''''
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|-
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| Trial||Luminol||H<sub>2</sub>O<sub>2</sub>||4-Iodophenol||HRP||Water||0.833mM Sodium Carbonate Buffer
 +
|-
 +
| 1||454 µL at 0.564 mM||112.7 µL at 5 mM||13.2 µL at 18 mM||33 µL at 9.6 µM||1033 µL||x
 +
|-
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| 2||454 µL at 0.564 mM||112.7 µL at 5 mM||13.2 µL at 18 mM||33 µL at 9.6 µM||1033 µL||x
 +
|-
 +
| 3||454 µL at 0.564 mM||112.7 µL at 5 mM||13.2 µL at 18 mM||33 µL at 9.6 µM||2000 µL||x
 +
|-
 +
| 4||454 µL at 0.564 mM||112.7 µL at 5 mM||13.2 µL at 18 mM||66 µL at 0.96 µM||2000 µL||x
 +
|-
 +
| 5||454 µL at 0.564 mM||112.7 µL at 5 mM||13.2 µL at 18 mM||33 µL at 2.3 µM||x||1000 µL
 +
|-
 +
| 6||454 µL at 0.564 mM||112.7 µL at 5 mM||13.2 µL at 18 mM||16.5 µL at 2.3 µM||x||1000 µL
 +
|-
 +
| 7||454 µL at 0.564 mM||112.7 µL at 5 mM||13.2 µL at 18 mM||16.5 µL at 0.96 µM||x||1000 µL
 +
|-
 +
| 8||454 µL at 0.564 mM||112.7 µL at 5 mM||13.2 µL at 18 mM||16.5 µL at 2.3 µM||x||2000 µL
 +
|}
  
 
==Data==
 
==Data==

Revision as of 21:53, 6 December 2012

<!-- sibboleth --><div id="lncal1" style="border:0px;"><div style="display:none;" id="id">lncal1</div><div style="display:none;" id="dtext"></div><div style="display:none;" id="page">User:Dhea Patel/Notebook/Experimental Biological Chemistry Notebook/2012/10/24</div><div style="display:none;" id="fmt">yyyy/MM/dd</div><div style="display:none;" id="css">OWWNB</div><div style="display:none;" id="month"></div><div style="display:none;" id="year"></div><div style="display:none;" id="readonly">Y</div></div>

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Objective

  • to run Fluorescence on Luminol-HRP assays.

Description

Please refer to table for volumes and concentrations of HRP, Luminol, Phenol, Buffer, and hydrogen peroxide used.

' Volume and Stock Concentration Added to Cuvette ' ' ' Volume Added to Cuvette [µL] '
Trial Luminol H2O2 4-Iodophenol HRP Water 0.833mM Sodium Carbonate Buffer
1 454 µL at 0.564 mM 112.7 µL at 5 mM 13.2 µL at 18 mM 33 µL at 9.6 µM 1033 µL x
2 454 µL at 0.564 mM 112.7 µL at 5 mM 13.2 µL at 18 mM 33 µL at 9.6 µM 1033 µL x
3 454 µL at 0.564 mM 112.7 µL at 5 mM 13.2 µL at 18 mM 33 µL at 9.6 µM 2000 µL x
4 454 µL at 0.564 mM 112.7 µL at 5 mM 13.2 µL at 18 mM 66 µL at 0.96 µM 2000 µL x
5 454 µL at 0.564 mM 112.7 µL at 5 mM 13.2 µL at 18 mM 33 µL at 2.3 µM x 1000 µL
6 454 µL at 0.564 mM 112.7 µL at 5 mM 13.2 µL at 18 mM 16.5 µL at 2.3 µM x 1000 µL
7 454 µL at 0.564 mM 112.7 µL at 5 mM 13.2 µL at 18 mM 16.5 µL at 0.96 µM x 1000 µL
8 454 µL at 0.564 mM 112.7 µL at 5 mM 13.2 µL at 18 mM 16.5 µL at 2.3 µM x 2000 µL

Data

Trail 6.png

  • All the trails, except trial 6 had a straight peak, showing do drop off of the fluorescence. Trail 6, containing, 454uL 0.564mM Luminol, 112.7uL 5mM hydrogen peroxide, 16.5uL 0.9uM HRP, 13.2 uL 18mM 4-Iodophenol, and 1000uL Carbonate Buffer, showed a distinct drop off. Tests should be run, keeping the molarity of 0.9uM constant and varying other factors to determine the optimum molar ratio.