User:Dhea Patel/Notebook/Experimental Biological Chemistry Notebook/2012/10/24

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(Objective)
(Description)
Line 23: Line 23:
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.
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[[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||Carbonate Buffer
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|-
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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
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|-
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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
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|-
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| 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
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|-
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| 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
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|-
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| 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
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|-
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| 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
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|-
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| 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
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|-
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| 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 00:47, 7 December 2012

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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] '
TrialLuminolH2O24-IodophenolHRPWaterCarbonate Buffer
1454 µL at 0.564 mM112.7 µL at 5 mM13.2 µL at 18 mM33 µL at 9.6 µM1033 µLx
2454 µL at 0.564 mM112.7 µL at 5 mM13.2 µL at 18 mM33 µL at 9.6 µM1033 µLx
3454 µL at 0.564 mM112.7 µL at 5 mM13.2 µL at 18 mM33 µL at 9.6 µM2000 µLx
4454 µL at 0.564 mM112.7 µL at 5 mM13.2 µL at 18 mM66 µL at 0.96 µM2000 µLx
5454 µL at 0.564 mM112.7 µL at 5 mM13.2 µL at 18 mM33 µL at 2.3 µMx1000 µL
6454 µL at 0.564 mM112.7 µL at 5 mM13.2 µL at 18 mM16.5 µL at 2.3 µMx1000 µL
7454 µL at 0.564 mM112.7 µL at 5 mM13.2 µL at 18 mM16.5 µL at 0.96 µMx1000 µL
8454 µL at 0.564 mM112.7 µL at 5 mM13.2 µL at 18 mM16.5 µL at 2.3 µMx2000 µL

Data

Image: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.


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