User:Michael F. Nagle/Notebook/Chem 571/2012/10/02

(Difference between revisions)

Revision as of 01:10, 7 December 2012

Project name

Prepare reagents for horseradish peroxidase (HRP) assay
- Complete HRP assay with and without gold nanoparticles to see if they interact with HRP's active site

HRP procedure¹ was followed. Time was plotted against absorbance on the UV/Vis spectroscoper.
1mg horseradish peroxidase was dissolved in 1mL distilled water.
A .2M buffer of sodium phosphate was made at a pH of 7.
.0017M hydrogen peroxide solution was made by adding 1mL 30% hydrogen peroxide to 100mL distilled water. 1mL of hydrogen peroxide solution was diluted to 50mL with 49mL of sodium phosphate buffer for 34μM solution.
698μM iodophenol solution was made in 1mL DMSO, since it's not soluble in water. 2.5mM 4-aminoantipyrine solution was prepared with the sodium phosphate buffer.

The HRP assays were not completed today, but will be 10/02, with the reagents prepared today

1. Horseradish Peroxidase Assay. Gold Biotechnology Image:3760-Protocol HRP AbsorbanceAssay.pdf

@@ Line 12: / Line 12: @@
 ==Procedure==
 #HRP procedure<sup>1</sup> was followed. Time was plotted against absorbance on the UV/Vis spectroscoper.
-#1mL horseradish peroxidase was dissolved in 1mL distilled water.
+#1mg horseradish peroxidase was dissolved in 1mL distilled water.
 #A .2M buffer of sodium phosphate was made at a pH of 7.
 #.0017M hydrogen peroxide solution was made by adding 1mL 30% hydrogen peroxide to 100mL distilled water. 1mL of hydrogen peroxide solution was diluted to 50mL with 49mL of sodium phosphate buffer for 34μM solution.