Difference between revisions of "User:Matt Hartings/Notebook/Photosynthesis/2013/10/17"

From OpenWetWare
Jump to: navigation, search
(Procedure and Data)
(fix raw html notebook nav)
 
(2 intermediate revisions by one other user not shown)
Line 2: Line 2:
 
|-
 
|-
 
|style="background-color: #EEE"|[[Image:Hartings_AU_Photosynthesis_Lab_Header.png|128px]]<span style="font-size:22px;"> Protein Re-engineering</span>
 
|style="background-color: #EEE"|[[Image:Hartings_AU_Photosynthesis_Lab_Header.png|128px]]<span style="font-size:22px;"> Protein Re-engineering</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>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
+
|style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}}
 
|-
 
|-
 
| colspan="2"|
 
| colspan="2"|
Line 11: Line 11:
  
 
==Objective==
 
==Objective==
Take kinetic data for Mb (lyophilized from KCl/acetate buffer) catalyzed o-phenylenediamine + H2O2 reaction in methanol (45C). Take kinetic data for Mb (lyophilized from KCl/phosphate buffer) catalyzed o-phenylenediamine + H2O2 reaction in water (5C, 15C, and 25C).
+
Take kinetic data for Mb (lyophilized from KCl/acetate buffer) catalyzed o-phenylenediamine + H2O2 reaction in methanol (45C). Take kinetic data for Mb (lyophilized from KCl/phosphate buffer) catalyzed o-phenylenediamine + H2O2 reaction in methanol (5C, 15C, and 25C).
  
 
==Procedure and Data==
 
==Procedure and Data==
Line 56: Line 56:
 
### Stop after 2 hours
 
### Stop after 2 hours
 
## Files saved as: Mb_OPD_H2O2_phosphate_meoh_5_SCANNUMBER.txt
 
## Files saved as: Mb_OPD_H2O2_phosphate_meoh_5_SCANNUMBER.txt
 +
## 0.225mL Mb stock solution added just before the third scan
 +
## This is denoted in the data file for the third scan
 +
 +
<u>sample 3</u>
 +
# Stock solutions:
 +
## Myoglobin
 +
### No myoglobin was used in this sample
 +
## OPD
 +
### 3.6mg OPD in 3.6mL methanol
 +
## H<sub>2</sub>O<sub>2</sub>
 +
### 0.114mL 30% H<sub>2</sub>O<sub>2</sub> in 0.887mL methanol
 +
#Experimental Run
 +
## Sample chamber set to 5C and to stir
 +
## N<sub>2</sub> run through the cuvette holder to reduce condensation on cuvette
 +
## 2.025mL OPD stock solution
 +
## 0.75mL H<sub>2</sub>O<sub>2</sub> stock solution
 +
### Start Data Collection
 +
### 1ms Integration
 +
### 10 scan average
 +
### Wait 1 minute after each scan
 +
### Stop after 2 hours
 +
## Files saved as: OPD_H2O2_phosphate_meoh_5_SCANNUMBER.txt
 
## 0.225mL Mb stock solution added just before the third scan
 
## 0.225mL Mb stock solution added just before the third scan
 
## This is denoted in the data file for the third scan
 
## This is denoted in the data file for the third scan

Latest revision as of 23:29, 26 September 2017

Hartings AU Photosynthesis Lab Header.png Protein Re-engineering Report.pngMain project page
Resultset previous.pngPrevious entry      Next entryResultset next.png



Objective

Take kinetic data for Mb (lyophilized from KCl/acetate buffer) catalyzed o-phenylenediamine + H2O2 reaction in methanol (45C). Take kinetic data for Mb (lyophilized from KCl/phosphate buffer) catalyzed o-phenylenediamine + H2O2 reaction in methanol (5C, 15C, and 25C).

Procedure and Data

sample 1

  1. Stock solutions:
    1. Myoglobin
      1. 1.3mg (Mb:KCl lyophilized from Citrate, 5mg:500mg) in 1.0mL methanol
    2. OPD
      1. 3.6mg OPD in 3.6mL methanol
    3. H2O2
      1. 0.114mL 30% H2O2 in 0.887mL methanol
  2. Experimental Run
    1. Sample chamber set to 45C and to stir
    2. N2 run through the cuvette holder to reduce condensation on cuvette
    3. 2.025mL OPD stock solution
    4. 0.75mL H2O2 stock solution
      1. Start Data Collection
      2. 1ms Integration
      3. 10 scan average
      4. Wait 1 minute after each scan
      5. Stop after 2 hours
    5. Files saved as: Mb_OPD_H2O2_Acetate_meoh_45_SCANNUMBER.txt
    6. 0.225mL Mb stock solution added just before the third scan
    7. This is denoted in the data file for the third scan

sample 2

  1. Stock solutions:
    1. Myoglobin
      1. 1.4mg (Mb:KCl lyophilized from Citrate, 5mg:500mg) in 1.0mL methanol
    2. OPD
      1. 2.4mg OPD in 2.4mL methanol
    3. H2O2
      1. 0.114mL 30% H2O2 in 0.887mL methanol
  2. Experimental Run
    1. Sample chamber set to 5C and to stir
    2. N2 run through the cuvette holder to reduce condensation on cuvette
    3. 2.025mL OPD stock solution
    4. 0.75mL H2O2 stock solution
      1. Start Data Collection
      2. 1ms Integration
      3. 10 scan average
      4. Wait 1 minute after each scan
      5. Stop after 2 hours
    5. Files saved as: Mb_OPD_H2O2_phosphate_meoh_5_SCANNUMBER.txt
    6. 0.225mL Mb stock solution added just before the third scan
    7. This is denoted in the data file for the third scan

sample 3

  1. Stock solutions:
    1. Myoglobin
      1. No myoglobin was used in this sample
    2. OPD
      1. 3.6mg OPD in 3.6mL methanol
    3. H2O2
      1. 0.114mL 30% H2O2 in 0.887mL methanol
  2. Experimental Run
    1. Sample chamber set to 5C and to stir
    2. N2 run through the cuvette holder to reduce condensation on cuvette
    3. 2.025mL OPD stock solution
    4. 0.75mL H2O2 stock solution
      1. Start Data Collection
      2. 1ms Integration
      3. 10 scan average
      4. Wait 1 minute after each scan
      5. Stop after 2 hours
    5. Files saved as: OPD_H2O2_phosphate_meoh_5_SCANNUMBER.txt
    6. 0.225mL Mb stock solution added just before the third scan
    7. This is denoted in the data file for the third scan