TY - JOUR
T1 - Isolation of inositol 1,4,5-trisphosphate receptor-associating proteins and selective knockdown using RNA interference.
AU - Mizutani, Akihiro
AU - Kawaai, Katsuhiro
AU - Hisatsune, Chihiro
AU - Ando, Hideaki
AU - Michikawa, Takayuki
AU - Mikoshiba, Katsuhiko
PY - 2010
Y1 - 2010
N2 - 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.
AB - 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.
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U2 - 10.1007/978-1-60327-175-2_9
DO - 10.1007/978-1-60327-175-2_9
M3 - Article
C2 - 20645186
AN - SCOPUS:78049371827
VL - 645
SP - 133
EP - 141
JO - Methods in Molecular Biology
JF - Methods in Molecular Biology
SN - 1064-3745
ER -