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Team:Calgary/Notebook/Protocols/PrussianBlueFerritinPHOptimization
From 2013.igem.org
(Difference between revisions)
| Line 9: | Line 9: | ||
<h2>Reaction Mixture</h2> | <h2>Reaction Mixture</h2> | ||
<ul> | <ul> | ||
| - | <li> 20μL Prussian Blue Ferritin ( | + | <li> 20μL Prussian Blue Ferritin (22μg/mL) </li> |
<li> 16μL Substrate (TMB or ABTS, 10mg/mL) </li> | <li> 16μL Substrate (TMB or ABTS, 10mg/mL) </li> | ||
<li> 32μL Hydrogen Peroxide (30%) </li> | <li> 32μL Hydrogen Peroxide (30%) </li> | ||
| - | <li> Buffer (pH | + | <li> Buffer (pH 2-11) up to 268μL </li> |
</ul> | </ul> | ||
<br> | <br> | ||
| - | <p> The reaction was conducted in room temperature for 10 minutes and the absorbance value was recorded at 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. The experiment was conducted multiple times with buffers of | + | <p> The reaction was conducted in room temperature for 10 minutes and the absorbance value was recorded at 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. The experiment was conducted multiple times with buffers of the vary pHs of 2, 2.73, 3.6, 4, 4.4, 5, 5.6, 8, 9 and 11. </p> |
</section> | </section> | ||
</html> | </html> | ||
Revision as of 08:39, 27 September 2013
pH Optimization
pH Optimization
Reaction Mixture
- 20μL Prussian Blue Ferritin (22μg/mL)
- 16μL Substrate (TMB or ABTS, 10mg/mL)
- 32μL Hydrogen Peroxide (30%)
- Buffer (pH 2-11) up to 268μL
The reaction was conducted in room temperature for 10 minutes and the absorbance value was recorded at 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. The experiment was conducted multiple times with buffers of the vary pHs of 2, 2.73, 3.6, 4, 4.4, 5, 5.6, 8, 9 and 11.
