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

Current revision

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

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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-| Trial||Luminol||H<sub>2</sub>O<sub>2</sub>||4-Iodophenol||HRP||Water||Carbonate Buffer
+| 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