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Assay for detection of transient intracellular Ca2+
专利权人:
Rajiv Gandhi Centre for Biotechnology;Department of Biotechnology
发明人:
Ramakrishnapillai Omkumar Vyomakesannair,Raveendran Rajeev Kumar,Steephan Mathew,Madhavan Mayadevi,Sudarsana Devi Suma Priya,Chandrika Arunkumar Renganathan,Mohanan Archana Geetha
申请号:
US201213661787
公开号:
US9958462(B2)
申请日:
2012.10.26
申请国别(地区):
美国
年份:
2018
代理人:
The Webb Law Firm
摘要:
This invention relates to a simple end point assay for detection of transient intracellular Ca2+ with broad applicability to many Ca2+ channel proteins comprising, Generation of expression constructs for the fusion proteins having the Ca2+/calmodulin dependent protein kinase II (CaMKII) phosphorylation sites of NR2A or NR2B subunits of N-methyl-D-aspartate receptor (NMDAR) or the voltage gated potassium channel of Drosophila (Eag) or any protein sequence which binds to the T-site of CaMKII similar to NR2B, conjugated to mitochondrial localizing signal sequence, or mutants of these sequences as described herein. Generation of mammalian expression constructs of α-CaMKll as a chimera with green fluorescent protein (GFP-α-CaMKII) or its mutants as described herein. Site-Directed mutagenesis, Transfection, Ca2+ stimulation, imaging and quantification of the number of cells with Ca2+-dependent signal, wherein, NMDA receptor activity assay, TRPVI receptor activity assay, GluR4 receptor activity assay are performed
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