Assay for detection of transient intracellular CA2
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) phosphorylat...
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| creator | SUDARSANADEVI SUMA PRIYA RAVEENDRAN RAJEEV KUMAR MADHAVAN MAYADEVI STEEPHAN MATHEW OMKUMAR VYOMAKESANNAIR RAMAKRISHNAPILLAI |
| description | 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. |
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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.</description><language>eng</language><subject>INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES ; MEASURING ; PHYSICS ; TESTING</subject><creationdate>2012</creationdate><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://worldwide.espacenet.com/publicationDetails/biblio?FT=D&date=20121106&DB=EPODOC&CC=US&NR=8304198B2$$EHTML$$P50$$Gepo$$Hfree_for_read</linktohtml><link.rule.ids>230,308,777,882,25545,76296</link.rule.ids><linktorsrc>$$Uhttps://worldwide.espacenet.com/publicationDetails/biblio?FT=D&date=20121106&DB=EPODOC&CC=US&NR=8304198B2$$EView_record_in_European_Patent_Office$$FView_record_in_$$GEuropean_Patent_Office$$Hfree_for_read</linktorsrc></links><search><creatorcontrib>SUDARSANADEVI SUMA PRIYA</creatorcontrib><creatorcontrib>RAVEENDRAN RAJEEV KUMAR</creatorcontrib><creatorcontrib>MADHAVAN MAYADEVI</creatorcontrib><creatorcontrib>STEEPHAN MATHEW</creatorcontrib><creatorcontrib>OMKUMAR VYOMAKESANNAIR RAMAKRISHNAPILLAI</creatorcontrib><title>Assay for detection of transient intracellular CA2</title><description>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. 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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.</abstract><oa>free_for_read</oa></addata></record> |
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| title | Assay for detection of transient intracellular CA2 |
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