Peroxidase UV Vis assay - Revision history

Jwelker at 19:58, 30 June 2020

2020-06-30T19:58:14Z

Jwelker at 19:57, 30 June 2020

2020-06-30T19:57:41Z

Jwelker: /* Progress as of December 2018 */

2018-11-28T05:15:07Z

Progress as of December 2018

Jwelker: /* Progress as of December 2018 */

2018-11-28T05:08:59Z

Progress as of December 2018

Jwelker at 04:58, 28 November 2018

2018-11-28T04:58:39Z

Jwelker at 04:58, 28 November 2018

2018-11-28T04:58:07Z

Bes at 14:55, 13 November 2018

2018-11-13T14:55:05Z

Jwelker at 01:46, 12 November 2018

2018-11-12T01:46:24Z

Jwelker: /* Standard Mixing Procedure */

2018-11-12T01:41:55Z

Standard Mixing Procedure

Jwelker: /* Standard Mixing Procedure */

2018-11-12T01:41:32Z

Standard Mixing Procedure

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 The desired amount of peroxidase is then added to the assay before the UV-Vis is ran.
 ==References==
 [[Media:J._Biol._Chem.-1984-Deits-13525-33.pdf|Purification and Assay publication]]
 ==Progress as of December 2018==

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-The focus of our research is the activity of sea urchin ovoperoxidase. We are starting with Horse Radish Peroxidase (HRP) to develop a standard protocol for measuring the activity of a peroxidase.
+This research project involved the oxidation of guaiacol using the reaction between horseradish peroxidase and hydrogen peroxide. The purpose of this project was to develop a protocol for measuring the activity of a peroxidase using the UV-Vis.
 ==Standard Mixing Procedure==
 The mixture of Peroxidase UV-Vis assays requires the use of a Guaiacol stock solution and a hydrogen peroxide stock solution.

← Older revision		Revision as of 05:15, 28 November 2018
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	The main focus of the first semester of this research project has been to develop an efficient lab protocol for measuring the activity of a peroxidase. Since we will eventually be working with sea urchin peroxidase, background research was done on how to properly care for sea urchins as well as how to make UV-Vis assays for sea urchin peroxidase. With this information, we developed a standard mixing protocol for the UV-Vis assay using HRP. Several weeks have been spent modifying this procedure to produce more consistent results. Brad has taught me how to use the kinetics function for the UV-Vis, how to analyze data using Igor, and how to use the mixing attachment for the UV-Vis. We have also set up the tank which will hold our sea urchins when they arrive next semester.		The main focus of the first semester of this research project has been to develop an efficient lab protocol for measuring the activity of a peroxidase. Since we will eventually be working with sea urchin peroxidase, background research was done on how to properly care for sea urchins as well as how to make UV-Vis assays for sea urchin peroxidase. With this information, we developed a standard mixing protocol for the UV-Vis assay using HRP. Several weeks have been spent modifying this procedure to produce more consistent results. Brad has taught me how to use the kinetics function for the UV-Vis, how to analyze data using Igor, and how to use the mixing attachment for the UV-Vis. We have also set up the tank which will hold our sea urchins when they arrive next semester.
		+
		+	==Spring Semester 2019 Goals==
		+
		+	Moving forward, we hope to establish a final lab protocol for measuring peroxidase. The beginning of the semester will focus on adjusting the sea urchins to their tank and effectively monitoring their environment. We will be collaborating with Laura Sanchez to synthesize the sea urchin ovoperoxidase.

← Older revision		Revision as of 05:08, 28 November 2018
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	==Progress as of December 2018==		==Progress as of December 2018==
		+
		+	The main focus of the first semester of this research project has been to develop an efficient lab protocol for measuring the activity of a peroxidase. Since we will eventually be working with sea urchin peroxidase, background research was done on how to properly care for sea urchins as well as how to make UV-Vis assays for sea urchin peroxidase. With this information, we developed a standard mixing protocol for the UV-Vis assay using HRP. Several weeks have been spent modifying this procedure to produce more consistent results. Brad has taught me how to use the kinetics function for the UV-Vis, how to analyze data using Igor, and how to use the mixing attachment for the UV-Vis. We have also set up the tank which will hold our sea urchins when they arrive next semester.

← Older revision		Revision as of 04:58, 28 November 2018
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	==Hi-Tech Mixing Procedure==		==Hi-Tech Mixing Procedure==
−
−	~~==Progress as of December 2018==~~


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	==Example Data==		==Example Data==
		+
		+
		+
		+	==Progress as of December 2018==

← Older revision		Revision as of 14:55, 13 November 2018
Line 24:		Line 24:

	==Hi-Tech Mixing Procedure==		==Hi-Tech Mixing Procedure==
		+
		+	==References==
		+	[[Media:J._Biol._Chem.-1984-Deits-13525-33.pdf\|Purification and Assay publication]]

	==Example Data==		==Example Data==

@@ Line 5: / Line 5: @@
 '''Guaiacol Stock Solution'''
 .5 microliters of Guaiacol is added to a 25 mL pH 8.0 buffer solution, which is comprised of 25 mL of DI water and a 8.0 pH buffer tablet that has been mixed thoroughly.
 '''Hydrogen Peroxide Stock Solution'''
 microliters of hydrogen peroxide is added to 1 mL of DI water.
 '''Assay'''
 .0 mL Guaiacol stock solution is added to a vial.
 microliters of the hydrogen peroxide stock solution is added to the Guaiacol stock solution.
-    The desired amount of peroxidase is then added to the assay before the UV-Vis is ran.
+The desired amount of peroxidase is then added to the assay before the UV-Vis is ran.
 ==Example Data==