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AAT Bioquest/Screen Quest™ CHO-Ga16 Chimera Cell line/38105/Each
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OverviewPrinterFriendlyVersionEx/Em(nm)None/NoneMWN/ACAS#N/ASolventN/AStorageN2(L)CategoryGPCRcAMPGPCRAssaysRelatedScreenQuest™celllinesareaseriesofcellsthathavebeensuccessfullyusedindrugdiscoveryandscreeningenvironmentsforstudyingG-protein-coupledreceptors(GPCR)thatdonotconventionallycouplethroughintracellularcalcium.IthavebeeneffectivelyusedwiththeFLIPR,FDSSSystemsinconjunctionwithnon-Gqcoupledmembersofmanyreceptorssuchaschemokine,SEROtonin,glutamate,dopamine,opoid,vasopressinand?-andß-adrenoceptorreceptorfamilies.Over60%oftheknownG-protein-coupledreceptorssignalthroughpathwaysotherthanGqwhichleadtoanincreaseinintracellularcalcium,andasgenomicsrevealsmoreG-protein-coupledreceptortargetsthistrendcontinuestoincrease.ScreenQuest™celllinesareusedforinvestigatingG-protein-coupledreceptors(GPCR)thatdonotconventionallycouplethroughintracellularcalcium.ScreenQuest™celllinesarebasedonaseriesofG-proteinchimeras,includingthepromiscuousG-protein,G?16.ThechimerasconsistofthealphasubunitofaGq-proteincomplexwhose5carboxy-terminalaminoacidshavebeenreplacedwiththosefromoneoftheotherG-proteins(eitherG?s,G?i,G?o,orG?z).TheseaminoacidsareresponsIBLeforthecouplingofthereceptortoitsG-protein.Co-expressionofthesechimeraswithspecificnon-Gq-coupledreceptorswhichnormallyactthroughthecAMPpathwaymayresultinthegenerationofanintracellularcalciumsignaluponreceptorstimulation.ScreenQuest™CHO-G?16celllineisCHO-K1cellsstablytransfectedwiththepromiscuousG-protein,G?16.WhenusedasahostcellfortransfectionexpressionofGiorGs-coupledreceptors,theconstitutivelyexpressedG?16proteininthecellsallowsthetransfectedreceptorwhichnormallyactthroughthecAMPpathway,tocoupletoGqsignaltransductionandmobilizedintracellularcalcium.Activationofthespecificnon-Gq-coupledreceptorsinthesecellsbyspecificligandscanbedetectedusingcalciumsensitivedyessuchasCal-520AM,Fluo-8AM,orFluo-4AM.Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds. ol { counter-reset: item; } ol > li { counter-increment: item; } ol ol > li { display: block; } ol ol > li:before { content: counters(item, ".") ". "; margin-left: -35px; } CellDensityandStorageThecellsarefrozenatadensityof2X106cellsin90%fetalbovineserumand10%(v/v)DMSO.ToinsurethehighestlevelofviABIlity,thawthevialandinitiatethecultureassoonaspossibleuponreceipt.Cellsmustbestoredinliquidnitrogenifnotimmediatelyprocesseduponreceipt.CellCultureMediumBasalmedium:Ham’sF12culturemediumcontaining10%fetalbovineserum,2mML-glutamine,100units/mLpenicillin,100µg/mLstreptomycin(Basalmedium).SelectionMedium:BasalMediumwith200µg/mLhygromycinBand400ug/mLG418.ThawingandSeedingCellsPrepareBasalMedium.Prepare37°CWaterBath.Thawcellsrapidlybyremovingfromliquidnitrogen,andimmediatelyimmersingina37°Cwaterbath.Removethevialfromthewaterbathassoonasthecontentsarethawed,andsterilizetheexteriorofthevialwith70%ethanol.Alloftheoperationsfromthispointonshouldbecarriedoutunderstrictasepticconditions.Addvialcontentsto25mLcellculturemediuminT75TissueCultureTreatedFlask.Gentlyswirlflaskandplaceinahumidified,tissuecultureincubator,37°C,5%CO2.Alllivecellsshouldbeattachedafter18-24hourspost–thaw.ChangeBasalMediumwithSelectionMedium.SubcultureandPropagationWhencellsareapproximately80%confluent,passagethecells.Passagethecells1:10every3-4days,usingTrypsin-EDTAtodissociatethecells.Itishighlyrecommendedthatafrozencellbankbeestablishedatlowpassagenumber. #referenceList a{color:#0259A2; font-size:14px;} #referenceList {color:#222;font-size:12px;} References&CitationsCitationExplorerActivationofP2X7andP2Y11purinergicreceptorsinhibitsmigrationandnormalizestumor-derivedendothelialcellsviacAMPsignalingAuthors:DAvanzato,TGenova,AFiorioPla,MBernardini,SBianco,BBussolati,DMancardi,EGiraudo,FMaione,PCassoniJournal:ScientificReports(2016)TheM2muscarinicreceptorsareessentialforsignalingintheheartleftventricleduringrestraintstressinmiceAuthors:HanaTomankova,PaulinaValuskova,EvaVarejkova,JanaRotkova,JanBenes,JaromirMyslivecekJournal:Stress(2015)
美国AAT Bioquest Inc.(前身是ABD Bioquest,Inc.)是一家为从事生命科学研究、诊断研发及药物开发的科学家研发、生产和销售生物分析研究试剂和试剂盒的公司。公司致力于光谱学检测领域,包括吸收(颜色)、荧光和发光技术。AAT Bioquest的产品帮助全世界的科学家和生物医药研究者更好的了解生物化学、免疫学、细胞生物学和分子生物学等领域。AAT Bioquest会不断研发新产品,快速地丰富各个领域的产品。
1)提供反应探针和发光探针,生物素和端粒酶能够应用于标记药物小分子和生物聚合物,如蛋白、核酸以及其他碳水化合物;
2)研究并生产荧光和发光探针用于检测蛋白,核酸和活细胞;
3)不断推出新型的荧光和发光探针用于检测多种酶,特别是检测水解酶和氧化还原酶类;
4)致力于开发用于信号转导研究的试剂;
5)提供生理和神经探针,特别是钙离子指示剂和膜电位探针。
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