Team:Calgary/Notebook/Protocols/PrussianBlueFerritinSynthesis

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<li>Horse Spleen Ferritin (47mg/mL) </li>
<li>Horse Spleen Ferritin (47mg/mL) </li>
<li>Hydrogen Chloride (pH 3.0)</li>
<li>Hydrogen Chloride (pH 3.0)</li>
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<li> K<sub>4</sub>Fe(CN)<sub>6</sub> (0.025M) </li>
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<li>K<sub>4</sub>Fe(CN)<sub>6</sub> (0.025M) </li>
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<li> Sodium acetate-acetic acid buffer (pH 3.6)</li>
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<li>Sodium acetate-acetic acid buffer (pH 3.6)</li>
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<p> 20µL of horsespleen ferritin (47mg/mL) was added to 20mL of HCl (pH 3.0) and 150µL of K<sub>4</sub>Fe(CN)<sub>6</sub> (0.025M). The solution was kept under 4℃ for 12 hours while constantly being stirred.</p>
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<li>Add 20µL horsespleen ferritin (47mg/mL) to a solution of 20mL HCl(pH 3.0) and 150µL K<sub>4</sub>Fe(CN)<sub>6</sub> (0.025M).</li>
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<p> After the 12 hour period, 5mL of the solution was dialyzed against 500mL of sodium acetate-acetic acid buffer (pH 3.6) for 24 hours, with the buffer being replaced after 8 hours.
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<li>Dialyze for 12 hours at 4℃ while constantly stirring</li>
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<li>Dialyze 5mL of the previous ferritin solution against sodium acetate-acetic acid buffer (pH 3.6) for 8 hours.</li>
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<li>Replace buffer and dialyze overnight </li>
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Revision as of 00:52, 28 September 2013

Prussian Blue Ferritin Synthesis

Reagents and Materials

  • Horse Spleen Ferritin (47mg/mL)
  • Hydrogen Chloride (pH 3.0)
  • K4Fe(CN)6 (0.025M)
  • Sodium acetate-acetic acid buffer (pH 3.6)

  1. Add 20µL horsespleen ferritin (47mg/mL) to a solution of 20mL HCl(pH 3.0) and 150µL K4Fe(CN)6 (0.025M).
  2. Dialyze for 12 hours at 4℃ while constantly stirring
  3. Dialyze 5mL of the previous ferritin solution against sodium acetate-acetic acid buffer (pH 3.6) for 8 hours.
  4. Replace buffer and dialyze overnight

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