User:Moira M. Esson/Notebook/CHEM-571/2013/09/17

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
(General Protocol)
(HRP Oxidation)
Line 10: Line 10:
<br>
<br>
==HRP Oxidation==
==HRP Oxidation==
-
<br>
+
General Protocol:
-
'''General Protocol''':
+
#Run UV-vis of a blank buffer sample.
#Run UV-vis of a blank buffer sample.
#Prepare a 1mL sample of 10μM HRP in 50Tris, 50Nacl, pH=7.5 buffer. This buffer should be degassed) directly in a clean glass cuvette.  
#Prepare a 1mL sample of 10μM HRP in 50Tris, 50Nacl, pH=7.5 buffer. This buffer should be degassed) directly in a clean glass cuvette.  

Revision as of 13:17, 17 September 2013

Project name Main project page
Previous entry      Next entry

Objectives

  1. Find endpoints (100%) of oxidation and reduction of HRP


HRP Oxidation

General Protocol:

  1. Run UV-vis of a blank buffer sample.
  2. Prepare a 1mL sample of 10μM HRP in 50Tris, 50Nacl, pH=7.5 buffer. This buffer should be degassed) directly in a clean glass cuvette.
  3. Take spectrum of the prepared 10μM HRP sample.
  4. Add 2μL of K3[Fe(CN)6] to the prepared sample.
  5. Take UV/vis spectrum of the sample.
  6. Repeat steps 4-5 until complete oxidation of HRP.
    • Note: When preparing the sample add all components 'very slowly
    • Note: Allow all samples to sit for approximately 5 minutes before taking the spectrum
    • Note: No more than 20uL should be added of the K3[Fe(CN)6] to the sample.




Personal tools