| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 573/588 |
| MW | N/A |
| CAS# | N/A |
| Solvent | N/A |
| Storage | F/D/L |
| Category | GPCR CalciumGPCRAssays |
| Related | CalciumChannels pHandIonIndicators |
| Spectrum | AdvancedSpectrumViewer |
- Preparecells:
- Foradherentcells:Platecellsovernightingrowthmediumat40,000to80,000cells/well/100µLfora96-wellplateor10,000to20,000cells/well/25µLfora384-wellplate.
- Fornon-adherentcells:CentrifugethecellsfromtheculturemediumandthensUSPendthecellpelletincellgrowthmediumorHHBSat125,000to250,000cells/well/100µLfora96-wellpoly-Dlysineplateor30,000to60,000cells/well/25µLfora384-wellpoly-Dlysineplate.Centrifugetheplateat800rpmfor2minuteswithbrakeoffpriortotheexperiments.
Note:Eachcelllineshouldbeevaluatedontheindividualbasistodeterminetheoptimalcelldensityfortheintracellularcalciummobilization.
- PrepareCalbryte™590AMdye-loADIngsolution:
- Thawallthekitcomponentsatroomtemperaturebeforeuse.
- MakeCalbryte™590AMstocksolution:Add20µL(forCat#36200)or200µL(forCat#36201and36202)ofDMSOintothevialofCalbryte™590AM(ComponentA),andmixthemwell.
Note:20µLofCalbryte™590AMstocksolutionisenoughforoneplate.UnusedCalbryte™590AMstocksolutioncanbealiquotedandstoredat<-20=""oc=""for=""more=""than=""one=""month=""if=""the=""tubes=""are=""sealed=""tightly.=""protect=""from=""light=""and=""avoid=""repeated=""freeze-thaw=""> - Make1Xassaybuffer:Mix9mLofHHBS(ComponentC,notincludedinthekitCat#36202)with1mLof10XPluronicF127Plus(ComponentB),andmixthemwell.
- MakeCalbryte™590AMdye-loadingsolutionforonecellplate:Add20µLofCalbryte™590AMstocksolution(fromStep2.2)into10mLof1Xassaybuffer(fromStep2.3),andmixthemwell.Thisworkingsolutionisstableforatleast2hoursatroomtemperature.
- Runcalciumassay:
- Add100µL/well(96-wellplate)or25µL/well(384-wellplate)ofCalbryte™590AMdye-loadingsolution(fromStep2.4)intothecellplate.
- Incubatethedye-loadingplateinacellincubatorfor~60minutes,andthenincubatetheplateatroomtemperatureforanother15minutes.
Note1:Iftheassayrequires37oC,performtheexperimentimmediatelywithoutfurtherroomtemperatureincubation.
Note2:Ifthecellscanfunctionwellatroomtemperatureforlongertime,incubatethecellplateatroomtemperaturefor1hour. - PreparethecompoundplatewithHHBSoryourdesiredbuffer.
- RunthecalciumfluxassaybymonitoringthefluorescenceintensitywithTRITCFilter(orEx/Em=540/590nm).
| References&Citations |  CitationExplorer |
CalreticulinregulatesTGF-β1-inducedepithelialmesenchymaltransitionthroughmodulatingSmadsignalingandcalciumsignaling
Authors:YanjiaoWu,XiaoliXu,LunkunMa,QianYi,WeichaoSun,LilingTang
Journal:TheInternationalJournalofBiochemistry&CellBIOLOGy(2017)
Dexmedetomidinereduceshypoxia/reoxygenationinjurybyregulatingmitochondrialfissioninrathippocampalneurons
Authors:JiaLiu,QingDu,HeZhu,YuLi,MaodongLiu,ShoushuiYu,ShileiWang
Journal:IntJClinExpMed(2017):6861--6868
Monosialoganglioside1mayalleviateneurotoxicityinducedbypropofolcombinedwithremifentanilinneuralstemcells
Authors:JiangLu,Xue-qinYao,XinLuo,YuWang,SookjaKimChung,He-xinTang,ChiWaiCheung,Xian-yuWang,ChenMeng,QingLi
Journal:NeuralRegenerationResearch(2017):945
Obtainingspontaneouslybeatingcardiomyocyte-likecellsfromadipose-derivedstromalvascularfractionsculturedonenzyme-crosslinkedgelatinhydrogels
Authors:GangYang,ZhenghuaXiao,XiaomeiRen,HaiyanLong,KunlongMa,HongQian,YingqiangGuo
Journal:ScientificReports(2017):41781
Di(2-ethylhexyl)phthalate-inducedapoptosisinratINS-1cellsisdependentonactivationofendoplasmicreticulumstressandsuppressionofantioxidantprotection
Authors:XiaSun,YiLin,QianshengHuang,JunpengShi,LingQiu,MeiKang,YajieChen,ChaoFang,TingYe,SijunDong
Journal:Journalofcellularandmolecularmedicine(2015):581--594
Theeffectofmitochondrialcalciumuniporteronmitochondrialfissioninhippocampuscellsischemia/reperfusioninjury
Authors:LantaoZhao,ShuhongLi,ShileiWang,NingYu,JiaLiu
Journal:Biochemicalandbiophysicalresearchcommunications(2015):537--542
FungusinducesthereleaseofIL-8inhumancornealepithelialcells,viaDectin-1-mediatedproteinkinaseCpathways.
Authors:Xu-DongPeng,Gui-QiuZhao,JingLin,NanJiang,QiangXu,Cheng-ChengZhu,Jain-QiuQu,LinCong,HuiLi
Journal:Internationaljournalofophthalmology(2014):441--447
Propofolandremifentanilatmoderateandhighconcentrationsaffectproliferationanddifferentiationofneuralstem/Progenitorcells
Authors:QingLi,JiangLu,XianyuWang
Journal:Neuralregenerationresearch(2014):2002
Roleofmitochondrialcalciumuniporterinregulatingmitochondrialfissioninthecerebralcortexesoflivingrats
Authors:NanLiang,PengWang,ShileiWang,ShuhongLi,YuLi,JinyingWang,MinWang
Journal:JournalofNeuralTransmission(2014):593--600
Increasedexpressionofcelladhesionmolecule1bymastcellsasacauseofenhancednerve--mastcellinteractioninahapten-inducedmousemodelofatopicdermatitis
Authors:MHagiyama,TInoue,TFuruno,TIino,SItami,MNakanishi,HAsada,YHosokawa,AIto
Journal:BritishJournalofDermatology(2013):771--778
AAT Bioquest AAT Bioquest是一家位于美国的生物公司，前身为ABD Bioquest，总部位于加利福尼亚州。专门从事光学检测技术十多年，一直致力于光谱学检测领域技术的创新和突破。其独特的光学检测技术，综合了化学、生物学和信息学等各个领域的研究，引领了比色、荧光和发光技术新一代光学探针的浪潮。AAT Bioquest在全球拥有强大的经验丰富的专业分销商网络，为从小型研究机构到《财富》500强企业的各类客户提供卓越的产品和定制服务。
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1）我们提供反应荧光探针和发光探针，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）我们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）我们不断的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）我们致力于开发用于信号转导研究的试剂；5）我们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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AATBioquest,Inc.(formerlyABDBioquest,Inc.)develops,manufacturesandmarketsbioanalyticalresearchreagentsandkitstoscientistsengagedinlifesciencesresearch,diagnosticR&Danddrugdiscovery.Wespecializeintheareaofphotometricdetectionsincludingabsorption(color),fluorescenceandluminescencetechnologies.TheCompany"sproductsenablescientistsandbiomedicalresearcherstobetterunderstandbiochemistry,immunology,cellBIOLOGyandmolecularbiology.AATBioquestconstantlyintroducesnewproducts,andoffersarapidlyexpandinglistofproductsthataregroupedintoseveralproductlines.
1)Ourreactivefluorescentandluminescentprobes,biotinsandtagenzymesareusedforlabelingsmalldrugmoleculesandbiopolymers,e.g,proteins,nucleicacidsandcarbohydrates;2)Wedevelopfluorescentandluminescentprobesfordetectingproteins,nucleicacidsandlivecells;3)Weconstantlyintroducenovelfluorescentandluminescentprobesfordetectingvariousenzymes,inparticular,hydrolyticandredoxenzymes;4)Wefocusondevelopingreagentsforsignaltransductionresearch;and5)Wealsoofferphysiologicalandneurologicalprobes,e.g.,calciumindicatorsandmembranepotentialprobes.
Besidesthestandardcatalogproductswealsooffercustomservicetomeetyourspecialresearchneeds.Ourcurrentservicesincludecustomsynthesisofcolorimetric,fluorescentandluminescentprobes,customdevelopmentofbiochemical,cell-basedanddiagnosticassaysandcustomscreeningofyourcompoundlibrariesagainstyourdefinedtargetsusingourvalidatedHTSassays.
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百奥森18分钟霉菌毒素检测试剂盒，有需要的可以了解下！多种规格型号可供选择
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牛胰岛素,是一种多肽
在1965年9月17日我国完成了结晶牛胰岛素的全合成。经过严格鉴定，它的结构、生物活力、物理化学性质、结晶形状都和天然的牛胰岛素完全一样。这是世界上第一个人工合成的蛋白质，为人类认识生命、揭开生命奥秘迈出了可喜的一大步。这项成果获1982年中国自然科学一等奖。
1953年，英国人F. SangerSanger由于测定了牛胰岛素的一级结构而获得1958年诺贝尔化学奖。
我用Promega公司的双荧光素酶检测试剂盒（E2920）检测到的firefly萤光素酶活性很低，只有4*10的3次方；海肾萤光素酶活性有10的6次方。
我用Ad293细胞做了转染，fireflyluciferase质粒：RanillaLuciferase质粒=0.1ug：0.025ug/一个孔（96孔板），共转染了二天，再进行双萤光素酶检测。
我想了解fireflyluciferase活性用promega的这个E2920-双萤光素酶试剂盒检测得到10的3次方，这种数值正常吗？
我做了3次重复实验，每次firefly萤光素酶活性很低，只有4*10的3次方；而海肾萤光素酶活性有10的6次方。
求指教？fireflyluciferase活性低？会是什么原因呢？
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