Isolation of Inositol 1,4,5-Trisphosphate Receptor-Associating Proteins and Selective Knockdown Using RNA Interference
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Inositol 1,4,5-trisphosphate (IP3 ) receptors (IP3 Rs) are IP3 -gated Ca2+ release channels localized on intracellular Ca2+ stores and play a role in the generation of complex patterns of intracellular Ca2+ signals. We show herein experimental protocols for the identification of associating proteins of IP3 R isoforms from various cells and tissues using affinity column chromatography and for the specific knockdown of the expression of IP3 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 Ca2+ signals.
