User:Moira M. Esson/Notebook/CHEM-571/2013/10/01

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#Buffer: 5mM Tris pH=8
#Buffer: 5mM Tris pH=8
#HRP: 1.7μM HRP in 50mL 5mM Tris pH=8  
#HRP: 1.7μM HRP in 50mL 5mM Tris pH=8  
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*Performed 20% dilution of 13.5mg luminol in 300μL DMSO added to 50mL 5mM Tris pH=8. (The luminol stock solution).
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*Performed 1/20 dilution of 13.5mg luminol in 300μL DMSO added to 50mL 5mM Tris pH=8. (The luminol stock solution).
-
**Make a 10mL stock solution of 20% dilute luminol stock solution.
+
**Make a 10mL stock solution of 1/20 dilute luminol stock solution.
 +
*Prepared stock solution of hydrogen peroxide was too dilute to measure (characteristic peak at 432 nm was not present)
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*Reaction did not occur because of the low concentration of hydrogen peroxide
   
   

Revision as of 14:19, 1 October 2013

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Objectives

  1. Monitor the kinetics and yield of horseradish peroxidase catalyzed oxidation of luminol
    1. Measure UV-Vis absorbance of luminol, HRP, and Luminol+HRP+H2O2
    2. Monitor chemiluminscence of luminol oxidation using stop-flow techniques
    3. Measure kinetics of reaction using stop-flow techniques and absorbance


UV-vis

Stock solutions prepared before class:

  1. Luminol: 13.5mg luminol in 300μL DMSO added to 50mL 5mM Tris pH=8
  2. Buffer: 5mM Tris pH=8
  3. HRP: 1.7μM HRP in 50mL 5mM Tris pH=8
  • Performed 1/20 dilution of 13.5mg luminol in 300μL DMSO added to 50mL 5mM Tris pH=8. (The luminol stock solution).
    • Make a 10mL stock solution of 1/20 dilute luminol stock solution.
  • Prepared stock solution of hydrogen peroxide was too dilute to measure (characteristic peak at 432 nm was not present)
  • Reaction did not occur because of the low concentration of hydrogen peroxide



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