User:Mary Mendoza/Notebook/CHEM572 Exp. Biological Chemistry II/2013/02/12 - Revision history

Mary Mendoza: /* Graphs */

Mary Mendoza — Wed, 08 May 2013 22:15:41 GMT

Graphs

←Older revision		Revision as of 22:15, 8 May 2013
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-	~~==Graphs==~~
-
-	~~[[Image:TableT1.JPG]]~~
-
-	~~[[Image:Avg_Velocity_T1.JPG]] [[Image:Lineweaver-Burk_Plot_T1.JPG]]~~

Mary Mendoza: /* ADA Kinetic Assay */

Mary Mendoza — Wed, 08 May 2013 22:15:27 GMT

ADA Kinetic Assay

←Older revision		Revision as of 22:15, 8 May 2013
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-	~~==ADA Kinetic Assay==~~
-	*The procedure was taken from [[User:Dhea Patel/Notebook/CHEM 572: ADA&Inhibitor Kinetics/2013/02/12\|Dhea Patel's]] notebook entry.
-	*The assay runs were continued from last week's, [[User:Mary Mendoza/Notebook/CHEM572 Exp. Biological Chemistry II/2013/02/05\|02/05/2013]], calculated values.
-	*The objective of performing the assay is to obtain the rate of the enzymatic reaction of adenosine into inosine by ADA.
-	*Baseline 200-400nm using pH 7.4, 0.05M phosphate buffer
-	*Temperature control: 25°C
-	*Phosphate buffer in reference cell
-	*Ran kinetics on 250uM solution (20uL ADA, 600uL 1250 uM adenosine, and 2.38mL pH 7.4, 0.05M phosphate buffer) for 5 minutes at 265 nm.
-	**absorbance exceeded 4.00, so the assay was stopped and a new solution was prepared.
-	*Ran kinetics on 2.56uM solution (20uL ADA, 600uL 12.8 uM adenosine, and 2.38mL pH 7.4, 0.05M phosphate buffer) for 5 minutes at 265 nm.
-	**dAbs was obtained and plotted vs. time in excel
-	*Ran kinetics on 6.4uM solution (20uL ADA, 600uL 32 uM adenosine, and 2.38mL pH 7.4, 0.05M phosphate buffer) for 5 minutes at 265 nm.
-	**dAbs was obtained and plotted vs. time in excel
-	*Ran kinetics on 16uM solution (20uL ADA, 600uL 80 uM adenosine, and 2.38mL pH 7.4, 0.05M phosphate buffer) for 5 minutes at 265 nm.
-	**dAbs was obtained and plotted vs. time in excel
-	*The following table describes the volumes and concentrations of the components added to the cuvette.

-
-	~~[[Image:Data_Table.JPG\|center]]~~
-
-
-	* Note: Since the 250 μM adenosine assay exhibited a high absorbance, this concentration was replaced with two other adenosine concentrations, 150 and 200 μM.
-	* The stock solution for 150 μM adenosine was made by diluting 150 μL of 5 mM adenosine into 850 ng/μL of 0.05 M phosphate buffer. The 200 μM adenosine was made by diluting 200 μL of 5 mM adenosine into 800 μL of 0.05 M phosphate buffer.

	==Graphs==		==Graphs==

Mary Mendoza: /* ADA Kinetic Assay */

Mary Mendoza — Wed, 13 Feb 2013 19:51:16 GMT

ADA Kinetic Assay

←Older revision		Revision as of 19:51, 13 February 2013
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	*Temperature control: 25°C		*Temperature control: 25°C
	*Phosphate buffer in reference cell		*Phosphate buffer in reference cell
-	*Ran kinetics on 250uM solution (20uL ADA, 600uL ~~1250uM Adenosine~~, and 2.38mL ~~pH7~~.4, 0.~~005M~~ phosphate buffer) for 5 minutes at ~~265nm~~.	+	*Ran kinetics on 250uM solution (20uL ADA, 600uL 1250 uM adenosine, and 2.38mL pH 7.4, 0.05M phosphate buffer) for 5 minutes at 265 nm.
	**absorbance exceeded 4.00, so the assay was stopped and a new solution was prepared.		**absorbance exceeded 4.00, so the assay was stopped and a new solution was prepared.
-	*Ran kinetics on 2.56uM solution (20uL ADA, 600uL 12.~~8uM Adenosine~~, and 2.38mL ~~pH7~~.4, 0.~~005M~~ phosphate buffer) for 5 minutes at ~~265nm~~.	+	*Ran kinetics on 2.56uM solution (20uL ADA, 600uL 12.8 uM adenosine, and 2.38mL pH 7.4, 0.05M phosphate buffer) for 5 minutes at 265 nm.
	**dAbs was obtained and plotted vs. time in excel		**dAbs was obtained and plotted vs. time in excel
-	*Ran kinetics on 6.4uM solution (20uL ADA, 600uL ~~32uM Adenosine~~, and 2.38mL ~~pH7~~.4, 0.~~005M~~ phosphate buffer) for 5 minutes at ~~265nm~~.	+	*Ran kinetics on 6.4uM solution (20uL ADA, 600uL 32 uM adenosine, and 2.38mL pH 7.4, 0.05M phosphate buffer) for 5 minutes at 265 nm.
	**dAbs was obtained and plotted vs. time in excel		**dAbs was obtained and plotted vs. time in excel
-	*Ran kinetics on 16uM solution (20uL ADA, 600uL ~~80uM Adenosine~~, and 2.38mL ~~pH7~~.4, 0.~~005M~~ phosphate buffer) for 5 minutes at ~~265nm~~.	+	*Ran kinetics on 16uM solution (20uL ADA, 600uL 80 uM adenosine, and 2.38mL pH 7.4, 0.05M phosphate buffer) for 5 minutes at 265 nm.
	**dAbs was obtained and plotted vs. time in excel		**dAbs was obtained and plotted vs. time in excel
	*The following table describes the volumes and concentrations of the components added to the cuvette.		*The following table describes the volumes and concentrations of the components added to the cuvette.

Mary Mendoza: /* ADA Kinetic Assay */

Mary Mendoza — Wed, 13 Feb 2013 18:08:23 GMT

ADA Kinetic Assay

←Older revision		Revision as of 18:08, 13 February 2013
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	* Note: Since the 250 μM adenosine assay exhibited a high absorbance, this concentration was replaced with two other adenosine concentrations, 150 and 200 μM.		* Note: Since the 250 μM adenosine assay exhibited a high absorbance, this concentration was replaced with two other adenosine concentrations, 150 and 200 μM.
	* The stock solution for 150 μM adenosine was made by diluting 150 μL of 5 mM adenosine into 850 ng/μL of 0.05 M phosphate buffer. The 200 μM adenosine was made by diluting 200 μL of 5 mM adenosine into 800 μL of 0.05 M phosphate buffer.		* The stock solution for 150 μM adenosine was made by diluting 150 μL of 5 mM adenosine into 850 ng/μL of 0.05 M phosphate buffer. The 200 μM adenosine was made by diluting 200 μL of 5 mM adenosine into 800 μL of 0.05 M phosphate buffer.
		+
		+	==Graphs==
		+
		+	[[Image:TableT1.JPG]]
		+
		+	[[Image:Avg_Velocity_T1.JPG]] [[Image:Lineweaver-Burk_Plot_T1.JPG]]
		+

	<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->		<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->

Mary Mendoza: /* ADA Kinetic Assay */

Mary Mendoza — Wed, 13 Feb 2013 18:02:58 GMT

ADA Kinetic Assay

←Older revision		Revision as of 18:02, 13 February 2013
Line 26:		Line 26:
	[[Image:Data_Table.JPG\|center]]		[[Image:Data_Table.JPG\|center]]

		+
		+	* Note: Since the 250 μM adenosine assay exhibited a high absorbance, this concentration was replaced with two other adenosine concentrations, 150 and 200 μM.
		+	* The stock solution for 150 μM adenosine was made by diluting 150 μL of 5 mM adenosine into 850 ng/μL of 0.05 M phosphate buffer. The 200 μM adenosine was made by diluting 200 μL of 5 mM adenosine into 800 μL of 0.05 M phosphate buffer.

	<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->		<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->

Mary Mendoza: /* ADA Kinetic Assay */

Mary Mendoza — Wed, 13 Feb 2013 05:14:41 GMT

ADA Kinetic Assay

←Older revision		Revision as of 05:14, 13 February 2013
Line 7:		Line 7:
	<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->		<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->
	==ADA Kinetic Assay==		==ADA Kinetic Assay==
		+	*The procedure was taken from [[User:Dhea Patel/Notebook/CHEM 572: ADA&Inhibitor Kinetics/2013/02/12\|Dhea Patel's]] notebook entry.
	*The assay runs were continued from last week's, [[User:Mary Mendoza/Notebook/CHEM572 Exp. Biological Chemistry II/2013/02/05\|02/05/2013]], calculated values.		*The assay runs were continued from last week's, [[User:Mary Mendoza/Notebook/CHEM572 Exp. Biological Chemistry II/2013/02/05\|02/05/2013]], calculated values.
	*The objective of performing the assay is to obtain the rate of the enzymatic reaction of adenosine into inosine by ADA.		*The objective of performing the assay is to obtain the rate of the enzymatic reaction of adenosine into inosine by ADA.

Mary Mendoza: /* ADA Kinetic Assay */

Mary Mendoza — Wed, 13 Feb 2013 05:09:32 GMT

ADA Kinetic Assay

←Older revision		Revision as of 05:09, 13 February 2013
Line 7:		Line 7:
	<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->		<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->
	==ADA Kinetic Assay==		==ADA Kinetic Assay==
-	*The assay runs were continued from last week's, [[User:Mary Mendoza/Notebook/CHEM572 Exp. Biological Chemistry II/2013/02/05\|02/05/2013]] calculated values.	+	*The assay runs were continued from last week's, [[User:Mary Mendoza/Notebook/CHEM572 Exp. Biological Chemistry II/2013/02/05\|02/05/2013]], calculated values.
	*The objective of performing the assay is to obtain the rate of the enzymatic reaction of adenosine into inosine by ADA.		*The objective of performing the assay is to obtain the rate of the enzymatic reaction of adenosine into inosine by ADA.
	*Baseline 200-400nm using pH 7.4, 0.05M phosphate buffer		*Baseline 200-400nm using pH 7.4, 0.05M phosphate buffer

Mary Mendoza: /* Entry title */

Mary Mendoza — Wed, 13 Feb 2013 05:08:55 GMT

Entry title

←Older revision		Revision as of 05:08, 13 February 2013
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	<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->		<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->
-	==~~Entry title~~==	+	==ADA Kinetic Assay==
-	* ~~Insert content here~~...	+	*The assay runs were continued from last week's, [[User:Mary Mendoza/Notebook/CHEM572 Exp. Biological Chemistry II/2013/02/05\|02/05/2013]] calculated values.
		+	*The objective of performing the assay is to obtain the rate of the enzymatic reaction of adenosine into inosine by ADA.
		+	*Baseline 200-400nm using pH 7.4, 0.05M phosphate buffer
		+	*Temperature control: 25°C
		+	*Phosphate buffer in reference cell
		+	*Ran kinetics on 250uM solution (20uL ADA, 600uL 1250uM Adenosine, and 2.38mL pH7.4, 0.005M phosphate buffer) for 5 minutes at 265nm.
		+	**absorbance exceeded 4.00, so the assay was stopped and a new solution was prepared.
		+	*Ran kinetics on 2.56uM solution (20uL ADA, 600uL 12.8uM Adenosine, and 2.38mL pH7.4, 0.005M phosphate buffer) for 5 minutes at 265nm.
		+	**dAbs was obtained and plotted vs. time in excel
		+	*Ran kinetics on 6.4uM solution (20uL ADA, 600uL 32uM Adenosine, and 2.38mL pH7.4, 0.005M phosphate buffer) for 5 minutes at 265nm.
		+	**dAbs was obtained and plotted vs. time in excel
		+	*Ran kinetics on 16uM solution (20uL ADA, 600uL 80uM Adenosine, and 2.38mL pH7.4, 0.005M phosphate buffer) for 5 minutes at 265nm.
		+	**dAbs was obtained and plotted vs. time in excel
		+	*The following table describes the volumes and concentrations of the components added to the cuvette.
		+
		+
		+	[[Image:Data_Table.JPG\|center]]

Mary Mendoza: Autocreate 2013/02/12 Entry for User:Mary_Mendoza/Notebook/CHEM572_Exp._Biological_Chemistry_II

Mary Mendoza — Wed, 13 Feb 2013 04:54:17 GMT

Autocreate 2013/02/12 Entry for User:Mary_Mendoza/Notebook/CHEM572_Exp._Biological_Chemistry_II

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{|{{table}} width="800"
|-
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>      </html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
|-
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==Entry title==
* Insert content here...


|}

__NOTOC__