Invention Grant
- Patent Title: Assay for detection of transient intracellular Ca2+
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Application No.: US13661787Application Date: 2012-10-26
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Publication No.: US09958462B2Publication Date: 2018-05-01
- Inventor: Omkumar Vyomakesannair Ramakrishnapillai , Rajeev Kumar Raveendran , Mathew Steephan , Mayadevi Madhavan , Suma Priya Sudarsana Devi , Arunkumar Renganathan Chandrika , Archana Geetha Mohanan
- Applicant: Rajiv Gandhi Centre for Biotechnology , Department of Biotechnology
- Applicant Address: IN Thiruvananthapuram IN New Delhi
- Assignee: Rajiv Gandhi Centre for Biotechnology,Department of Biotechnology
- Current Assignee: Rajiv Gandhi Centre for Biotechnology,Department of Biotechnology
- Current Assignee Address: IN Thiruvananthapuram IN New Delhi
- Agency: The Webb Law Firm
- Priority: IN1276/CHE/07 20070619
- Main IPC: C12N5/10
- IPC: C12N5/10 ; G01N33/84 ; G01N33/68 ; C07K14/435 ; C12N9/12 ; G01N33/50

Abstract:
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 to detect the activity of Ca2+ channel proteins.
Public/Granted literature
- US20130089868A1 Assay for Detection of Transient Intracellular CA2+ Public/Granted day:2013-04-11
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