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Gel Electrophoresis

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Chamodhi Ranathunga

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References

Analysis of Dictyostelium discoideum Inositol Pyrophosphate Metabolism by Gel Electrophoresis

  • Pisani, Francesca, Thomas Livermore, Giuseppina Rose, Jonathan Robert Chubb, Marco Gaspari, Adolfo Saiardi, and Thierry Soldati. "Analysis of Dictyostelium Discoideum Inositol Pyrophosphate Metabolism by Gel Electrophoresis." PLoS ONE (2014): E85533. Web. 8 Jan. 2015. <http://www.plosone.org/article/info:doi/10.1371/journal.pone.0085533>
  • "Biology Animation Library." DNALC Blogs. N.p., n.d. Web. 5 Jan. 2015.

Purpose:

  • PAGE technology vs. SaxHPLC
  • characterize Inositol Pyrophosphate metabolism by D. discoideum ameoba
  • Dictyostelium discoideum amoeba produces inositol pyrophosphate (IP) when controlling metabolism and insulin signalling.

Dictyostelium discoideum Amoeba

Gel Electrophoresis

Gel Electrophoresis

Procedure used to separate DNA fragments according to size

by: Chamodhi Ranathunga

How it works

Results

Characterization of D. discoideum IP5 species. (Fig 4)

Treatment by Phytase, Ddp1 and acidic degradation define IP6, IP7, and IP8, in D. discoideum cell extract (Fig 2)

Mass spectrometry analysis of inositol pyrophosphates purified from D. Discoideum cell extract. (Fig 3)

PAGE analysis of inositol pyrophosphate during D. Discoideum development. (Fig 5)

PAGE analysis of D. discodeum cell extract reveal presence of 3 major bands (Fig 1)

Methods used

No alteration of IP7 and IP8 metabolism after cAMP treatment. (Fig 6)

Conclusion

  • Sax-HPLC
  • (Strong Anion Exchange Chromatography)
  • PAGE
  • (Polyacrylamide Gel Electrophoresis)
  • Enzymology
  • study enzymes: confirm genuinty of IP
  • Spectrometery
  • peaks of IP7 and IP8
  • Used the technology to resolve and visualize inositol phosphates (which was to characterize in vitro enzymatic reaction)
  • Sax-HPLC method is inferior to that of PAGE
  • inexpensive
  • resolve different isomeric species
  • genetic system surpassing S. cerevisae model because of high complexity and greater similarity
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