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

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==Procedure==
==Procedure==
#HRP procedure [[Image:3760-Protocol HRP AbsorbanceAssay.pdf]] was followed
#HRP procedure [[Image:3760-Protocol HRP AbsorbanceAssay.pdf]] was followed. Time was plotted against absorbance on the UV/Vis spectroscoper.
#1mL horseradish peroxidase was dissolved in 1mL distilled water.  
#1mL horseradish peroxidase was dissolved in 1mL distilled water.  
#A .2M buffer of sodium phosphate was made at a pH of 7.
#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.  
#.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.
#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.
*'''[[User:Abigail E. Miller|Abigail E. Miller]] 11:46, 7 October 2012 (EDT)''':what is different? you did not follow it exactly. what mistakes did oyu make? how did you get to these solutions? everything must be included in your notebook!


==Discussion==
==Discussion==
#The HRP assays were not completed today, but will be [[User:Michael_F._Nagle/Notebook/Chem_571/2012/10/02|10/02]], with the reagents prepared today
#The HRP assays were not completed today, but will be [[User:Michael_F._Nagle/Notebook/Chem_571/2012/10/02|10/02]], with the reagents prepared today


*'''[[User:Abigail E. Miller|Abigail E. Miller]] 11:47, 7 October 2012 (EDT)''':you have data that you collected this day. it should be on this page. all data is relevant! you do not pick and chose what gets to go in your notebook.


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Revision as of 20:28, 25 October 2012

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Objective

  • Prepare reagents for HRP assay and complete HRP assay with and without gold nanoparticles to see how they affect the reaction.

Procedure

  1. HRP procedure File:3760-Protocol HRP AbsorbanceAssay.pdf was followed. Time was plotted against absorbance on the UV/Vis spectroscoper.
  2. 1mL horseradish peroxidase was dissolved in 1mL distilled water.
  3. A .2M buffer of sodium phosphate was made at a pH of 7.
  4. .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.
  5. 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.

Discussion

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



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