User:Dhea Patel/Notebook/Experimental Biological Chemistry Notebook/2012/09/19

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
(→Description)
Line 24: Line 24:
*10mL of 2.5mM Luminol stock was prepared using the following calculations:
*10mL of 2.5mM Luminol stock was prepared using the following calculations:
**'''MW luminol''': 177.16 g/mol
**'''MW luminol''': 177.16 g/mol
-
**'''Mass luminol''':
+
**'''Mass luminol''': 10 mL × (10<sup>-3</sup> L/mL) × (2.5 × 10<sup>-3</sup> M) × (10<sup>-3</sup> mol/mmol) × 177.16 g/mol = 0.00443 g luminol
-
        10 mL × (10<sup>-3</sup> L/mL) × (2.5 × 10<sup>-3</sup> M) × (10<sup>-3</sup> mol/mmol) × 177.16 g/mol = 0.00443 g luminol
+
**'''Actual mass luminol''': 0.0044 g
**'''Actual mass luminol''': 0.0044 g
-
**'''Actual concentration''':  
+
**'''Actual concentration''': 0.0044 g × (1 mol/177.16 g) × (1/0.010 L) = 2.5 × 10<sup>-3</sup> M in H<sub>2</sub>O
-
        0.0044 g × (1 mol/177.16 g) × (1/0.010 L) = 2.5 × 10<sup>-3</sup> M in H<sub>2</sub>O
+
***Because the luminol did not completely dissolve in H<sub>2</sub>O, 0.08g Na<sub>2</sub>CO<sub>3</sub> was added to aid in its dissolution.
***Because the luminol did not completely dissolve in H<sub>2</sub>O, 0.08g Na<sub>2</sub>CO<sub>3</sub> was added to aid in its dissolution.
****Ratio: 4g sodium carbonate/500mL solution
****Ratio: 4g sodium carbonate/500mL solution
***After the luminol dissolved with the sodium carbonate, 0.48g Na<sub>2</sub>HCO<sub>3</sub> was added to the solution to neutralize the pH of the solution to about pH 7.  
***After the luminol dissolved with the sodium carbonate, 0.48g Na<sub>2</sub>HCO<sub>3</sub> was added to the solution to neutralize the pH of the solution to about pH 7.  
-
*The first sample was mixed with 4-Iodophenol, H<sub>2</sub>O<sub>2</sub>, luminol, and HRP. Please refer to [User:Keyun_Wang/Notebook/Experimental_Biological_Chemistry_I/2012/09/19 Kay's entry] for details in initial concentration, final concentration, and volumes added.  
+
*The first sample was mixed with 4-Iodophenol, H<sub>2</sub>O<sub>2</sub>, luminol, and HRP. Please refer to [[User:Keyun_Wang/Notebook/Experimental_Biological_Chemistry_I/2012/09/19 Kay's entry]] for details in initial concentration, final concentration, and volumes added.  
<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->
<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->

Revision as of 13:23, 25 September 2012

Search this Project

Customize your entry pages

Objective

  1. To determine enzymatic activity of horseradish peroxidase via the usage of fluorometer under 510nm.

Description

  • The stock solutions of HRP, H2O2, and phenol that were made yesterday were mixed and run under the fluorometer at 510nm.
  • 10mL of 2.5mM Luminol stock was prepared using the following calculations:
    • MW luminol: 177.16 g/mol
    • Mass luminol: 10 mL × (10-3 L/mL) × (2.5 × 10-3 M) × (10-3 mol/mmol) × 177.16 g/mol = 0.00443 g luminol
    • Actual mass luminol: 0.0044 g
    • Actual concentration: 0.0044 g × (1 mol/177.16 g) × (1/0.010 L) = 2.5 × 10-3 M in H2O
      • Because the luminol did not completely dissolve in H2O, 0.08g Na2CO3 was added to aid in its dissolution.
        • Ratio: 4g sodium carbonate/500mL solution
      • After the luminol dissolved with the sodium carbonate, 0.48g Na2HCO3 was added to the solution to neutralize the pH of the solution to about pH 7.
  • The first sample was mixed with 4-Iodophenol, H2O2, luminol, and HRP. Please refer to User:Keyun_Wang/Notebook/Experimental_Biological_Chemistry_I/2012/09/19 Kay's entry for details in initial concentration, final concentration, and volumes added.


Retrieved from "http://www.openwetware.org/wiki/User:Dhea_Patel/Notebook/Experimental_Biological_Chemistry_Notebook/2012/09/19"
Personal tools