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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<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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<li>Incubate for 12 hours at 4℃ to allow the synthesis to occur.</li>
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<li>Dialyze the solution (or aliquots of the solution) using dialysis tubing and sodium acetate-acetic acid buffer (pH 3.6) for 8 hours. Replace the buffer and allow it to dialyze over night. The buffer used can be changed depending on the future uses planned for the Prussian blue ferritin. Dialysis specifications should be followed according to the manufacturer's instructions for the tubing.</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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<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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Latest revision as of 06:10, 23 October 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. Incubate for 12 hours at 4℃ to allow the synthesis to occur.
  3. Dialyze the solution (or aliquots of the solution) using dialysis tubing and sodium acetate-acetic acid buffer (pH 3.6) for 8 hours. Replace the buffer and allow it to dialyze over night. The buffer used can be changed depending on the future uses planned for the Prussian blue ferritin. Dialysis specifications should be followed according to the manufacturer's instructions for the tubing.

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