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

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(Description)
(Description)
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| align="center" style="background:#f0f0f0;"|''''''
 
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| Trial||Luminol||H<sub>2</sub>O<sub>2</sub>||4-Iodophenol||HRP||Water||Carbonate Buffer
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| Trial||Luminol||H<sub>2</sub>O<sub>2</sub>||4-Iodophenol||HRP||Water||0.833mM Sodium Carbonate Buffer
 
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|-
 
| 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
 
| 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

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.