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Team:Calgary/Notebook/Protocols/PrussianBlueFerritinTemperatureOptimization
From 2013.igem.org
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| Line 9: | Line 9: | ||
<h2>Reaction Mixture</h2> | <h2>Reaction Mixture</h2> | ||
<ul> | <ul> | ||
| - | <li> 10μL Prussian Blue Ferritin ( | + | <li> 10μL Prussian Blue Ferritin (22μg/mL) </li> |
<li> 6μL Substrate (TMB or ABTS, 10mg/mL) </li> | <li> 6μL Substrate (TMB or ABTS, 10mg/mL) </li> | ||
<li> 32μL Hydrogen Peroxide (30%) </li> | <li> 32μL Hydrogen Peroxide (30%) </li> | ||
Revision as of 08:40, 27 September 2013
Temperature Optimization
Prussian Blue Ferritin Temperature Optimization
Reaction Mixture
- 10μL Prussian Blue Ferritin (22μg/mL)
- 6μL Substrate (TMB or ABTS, 10mg/mL)
- 32μL Hydrogen Peroxide (30%)
- Sodium Acetate-Acetic Acid Buffer (pH 3.6) up to 242μL
The reaction was conducted at temperatures ranging from 25-45˚C, with 5˚C intervals. Reaction continued for 10 minutes and absorbance values were taken in 10 second intervals. Absorbance for TMB was taken at 650nm, and absorbance for ABTS was taken at 415nm. Eight replicates were conducted. Substrates were the last reagent to be added to the mix.
