Isolation of inositol 1,4,5-trisphosphate receptor-associating proteins and selective knockdown using RNA interference.

Akihiro Mizutani, Katsuhiro Kawaai, Chihiro Hisatsune, Hideaki Ando, Takayuki Michikawa, Katsuhiko Mikoshiba

Research output: Contribution to journalArticle

Abstract

Inositol 1,4,5-trisphosphate (IP(3)) receptors (IP(3)Rs) are IP(3)-gated Ca(2+) release channels localized on intracellular Ca(2+) stores and play a role in the generation of complex patterns of intracellular Ca(2+) signals. We show herein experimental protocols for the identification of associating proteins of IP(3)R isoforms from various cells and tissues using affinity column chromatography and for the specific knockdown of the expression of IP(3)R isoforms and their associating proteins using RNA interference. These methods will provide clues to understand the exact nature of how the signaling complex contributes to the generation of spatio-temporal patterns of intracellular Ca(2+) signals.

Original languageEnglish
Pages (from-to)133-141
Number of pages9
JournalMethods in molecular biology (Clifton, N.J.)
Volume645
DOIs
Publication statusPublished - 2010 Nov 8
Externally publishedYes

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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