发明名称 |
ASSAY FOR DETECTION OF TRANSIENT INTRACELLULAR CA2+ |
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-CaMKII 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 to detect the activity Of Ca2+ channel proteins. |
申请公布号 |
EP2162742(B1) |
申请公布日期 |
2015.09.30 |
申请号 |
EP20080789892 |
申请日期 |
2008.06.12 |
申请人 |
RAJIV GANDHI CENTRE FOR BIOTECHNOLOGY;DEPARTMENT OF BIOTECHNOLOGY |
发明人 |
OMKUMAR, RAMAKRISHNAPILLAI, VYOMAKESANNAIR;RAVEENDRAN, RAJEEV, KUMAR;STEEPHAN, MATHEW;MADHAVAN, MAYADEVI;SUDARSANADEVI, SUMA, PRIYA |
分类号 |
G01N33/68 |
主分类号 |
G01N33/68 |
代理机构 |
|
代理人 |
|
主权项 |
|
地址 |
|