User:Alexander Cvitan/Notebook/Experimental Biological Chemistry Lab/2013/10/16: Difference between revisions

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==Procedure==
==Procedure==
===Kinetics of HRP-AuNP's===
====Solutions Made====
* Luminol
** Stock was 1.46mM
** A 20 times dilution was made to make a final solution volume of .072mM. (This was done in order to observe an abs on UV-Vis less than 1).
** .15ml of Luminol was used for the reaction.
* Hydrogen Peroxide
**Stock was 44.29mM
** Was diluted to be 50 times the concentration of the Hydrogen Peroxide. Thus the concentration is, 3.65mM.
** .247ml of hydrogen peroxide was used for the reaction.
* HRP-AuNP's
** Originally 100ul of the 60:1 diluted sample was used. This solution was made using Au [] of .25mM and a HRP [] of 4.167uM. This caused an extremely slow kinetic reaction.
** This reaction was then repeated using 1ml of HRP-AuNP's. 
*For both kinetic runs solutions were added as dictated above and were filled to 3ml with buffer using a micropipete.


==Data==
==Data==
__NOTOC__
__NOTOC__

Revision as of 11:28, 16 October 2013

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Objective

  • To complete the procedure outlined by Dr.Hartings HERE.
  • Run UV-Vis on Citrate, BSA, HRP-Au nanoparticles. Run a water blank.
  • Run kinetic analysis on the 60:1 diluted HRP-AU nanoparticles.
  • Remake BSA nanoparticles. (The BSA nanoparticles made previously formed fibers, rendering them useless).

Procedure

Kinetics of HRP-AuNP's

Solutions Made

  • Luminol
    • Stock was 1.46mM
    • A 20 times dilution was made to make a final solution volume of .072mM. (This was done in order to observe an abs on UV-Vis less than 1).
    • .15ml of Luminol was used for the reaction.
  • Hydrogen Peroxide
    • Stock was 44.29mM
    • Was diluted to be 50 times the concentration of the Hydrogen Peroxide. Thus the concentration is, 3.65mM.
    • .247ml of hydrogen peroxide was used for the reaction.
  • HRP-AuNP's
    • Originally 100ul of the 60:1 diluted sample was used. This solution was made using Au [] of .25mM and a HRP [] of 4.167uM. This caused an extremely slow kinetic reaction.
    • This reaction was then repeated using 1ml of HRP-AuNP's.
  • For both kinetic runs solutions were added as dictated above and were filled to 3ml with buffer using a micropipete.

Data
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