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Inositol Phosphates Purification Using Titanium Dioxide Beads

2018; American Academy of Arts and Sciences; Volume: 8; Issue: 15 Linguagem: Inglês

10.21769/bioprotoc.2959

2331-8325

Miranda Wilson, Adolfo Saiardi,

Protease and Inhibitor Mechanisms

Inositol phosphates (IPs) comprise a family of ubiquitous eukaryotic signaling molecules. They have been linked to the regulation of a pleiotropy of important cellular activities, but low abundance and detection difficulties have hampered our understanding. Here we present a method to purify and enrich IPs or other phosphate-rich metabolites from mammalian cells or other sample types. Acid-extracted IPs from cells bind selectively via their phosphate groups to titanium dioxide beads. After washing, the IPs are easily eluted from the beads by increasing the pH. This technique, in combination with downstream analytical methods such as PAGE or SAX-HPLC, opens unprecedented investigative possibilities, allowing appropriate analysis of IPs from virtually any biological or non-biological source.