4000-520-616
欢迎来到免疫在线!(蚂蚁淘生物旗下平台)  请登录 |  免费注册 |  询价篮
aatbio(优势品牌)
主营:主营:研究并生产荧光和发光探针,信号转导研究的试剂
咨询热线电话
4000-520-616
当前位置: 首页 > 产品中心 > Other_kits > AAT Bioquest/Screen Quest™不洗钾通道分析试剂盒/36550/1平板
商品详细AAT Bioquest/Screen Quest™不洗钾通道分析试剂盒/36550/1平板
AAT Bioquest/Screen Quest™不洗钾通道分析试剂盒/36550/1平板
AAT Bioquest/Screen Quest™不洗钾通道分析试剂盒/36550/1平板
商品编号: 36550
品牌: aatbio
市场价: ¥56560.00
美元价: 33936.00
产地: 美国(厂家直采)
公司:
产品分类: 其它检测试剂盒
公司分类: Other_kits
联系Q Q: 3392242852
电话号码: 4000-520-616
电子邮箱: info@ebiomall.com
商品介绍
Overview
PrinterFriendlyVersion

Ex/Em(nm)490/525
MWN/A
CAS#N/A
SolventN/A
StorageF/D/L
CategoryIonChannels
Related
Potassium(K+)ionchannelplaysanimportantroleinregulatingfundamentalBIOLOGicalprocessesincludingheartrate,hormoneandneurotransmittersecretion,waterandelectrolytebalance.Potassiumionchannelhasbeenconsideredasdrugtargetsfordiseaseindicationsincludingarrhythmia,pain,diabetes,neurologicaldysfunctionsetc.ThepermeABIlityofTl+throughK+channelhasbeenwidelyusedtoassayK+channel.ThecellsthatexpressK+channelofinterests(e.g.hERG,Kv1.3,Kir2.1,KATP)arepre-loadedwithaTl+sensitivedye.Thedyeisnon-fluorescentandispermeabletocellmembrane.Onceinsidethecell,thenon-fluorescentAMesterdyeiscleavedbyendogenousesteraseintoanegativelychargesdyethatstaysinsidecells.WhenastimulusbuffercontaininglowdoseofTl+isaddedtocells,theTl+flowsacrosstheK+channelandbindstoTl+sensitivedye,generatingafluorescentsignal.ThissignalisproportionaltotheactivityofK+channel.IfanantagoNISTorantagonistisaddedtothecells,thefluorescentsignaldecreasesorincreasesrespectively,toreflecttheinhibitedorstimulatedactivityofK+channel.
SpectrumAdvancedSpectrumViewer

Sorry,yourbrowserdoesnotsupportinlineSVG.RelativeIntensity(%)100806040200Sorry,yourbrowserdoesnotsupportinlineSVG.
Sorry,yourbrowserdoesnotsupportinlineSVG.Sorry,yourbrowserdoesnotsupportinlineSVG.
Movemouseovergridtodisplaywavelength&intensityvalues.

300
400
500
600
700
800
900
Wavelength(nm)


Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.
  1. Preparecells:
    1. Foradherentcells:Platecellsovernightingrowthmediumat40,000to80,000cells/well/100µLfora96-wellpolyD-lysineplateor10,000to20,000cells/well/25µLfora384-wellpoly-D-lysineplate.
    2. Fornon-adherentcells:CentrifugethecellsfromtheculturemediumandthensUSPendthecellpelletinequalamountofHHBSandTl-520AMdye-loADIngsolution(seeStep2.3below)at125,000to250,000cells/well/100µLfora96-wellpoly-D-lysineplateor30,000to60,000cells/well/25µLfora384-wellpoly-D-lysineplate.Centrifugetheplateat800rpmfor2minuteswithbrakeoffpriortotheexperiments.
      Note1:Eachcelllineshouldbeevaluatedontheindividualbasistodeterminetheoptimalcelldensityfortheintracellularthalliummobilization.
      Note2:ForHEK293-KCNH2cellsthatweusedinthisassay,werecommendtoplatecellsat20,000cells/well/100µLina96-wellpoly-Dlysineplateforovernight.Thecellsshouldreach80-90%confluencyontheexperimentday.

  2. Preparedye-loadingsolution:
    1. Make500XTl-520AMstocksolution:Add20µL(forCat#36550)or200µL(forCat#36551and36552)ofDMSOintothevialofTl-520AM(ComponentA),andmixwell.
      Note:TheunusedTl-520AMstocksolutioncanbestoredat-20°Cinasingle-usealiquots.
    2. Make1Xassaybuffer:Add1mlof10XPluronic®F127Plus(ComponentB)to9mlofHHBS(ComponentC).
    3. Makedye-loadingsolution:Add20µLofdyestocksolution(fromStep2.2)into10mlof1Xassaybuffer(fromStep2.3),andmixwell.

  3. Incubatecellswithdye-loadingsolution:
    1. Addequalvolume(suchas100µL/wellfora96-wellplateor25µL/wellfora384-wellplate)ofDye-LoadingSolution(fromStep2.3)toyourcellplatewithoutremovingthecellmedium.
    2. Incubatethedye-ladingplateinthecellincubatorfor1hour.

  4. Prepare5Xstimulussolution:
    1. Make1Xchloridefreebuffer(ComponentD):Dilute5Xchloridefreebufferto1XchloridefreebufferinddH2O.
    2. MakeTl2SO4solution(Notprovided,SigmaCat#208191):DissolveTl2SO4inultrapureH2Otofinalconcentrationof80mM.
      Note:Tl2SO4istoxic.Takenecessaryprecautionstopreventinhalationandskincontact
    3. Preparea5XStimulussolutionbydilutingligands(fornon-voltagegatedpotassiumchannels)orK2SO4(forvoltagegatedpotassiumchannels)withTl2SO4in1XChloride-freebuffer.

      Pleaseusethefollowingtableasareference(forvoltagegatedhERGchannelinHEK293-KCNH2cells),concentrationofTl2SO4andK2SO4(forvoltagegatedpotassiumchannels)orligands(fornon-voltagegatedpotassiumchannels)usedfortheassayshouldbeoptimizedforeachtargetchannelandcelltype.

      ComponentVolume5XConcentrationFinalConcentration
      5XChloride-freeAssayBuffer(ComponentD)2mL
      250mMPotassiumSulfate(K2SO4)(ComponentE)1mL12.5mM2.5mM
      80mMThalliumSulfate(Tl2SO4)(notprovided)0.5mL2.5mM0.5mM
      ddH2O5.5mL
      Total:10mL

  5. Runthepotassiumassay:
    1. PreparetheK+channelantagonistsoragonist(from4.3)inHHBS.
    2. Incubatetheplatewithantagonists(forinhibitorystudy)inthecellincubatorfor30minutes.
    3. Add50µL/well(fora96-wellplate)or12.5µL/well(fora384-wellplate)of5XstimulusbufferwithFLIPR,FDSSorFlexstation.RuntheexperimentwithafiltersetofEx/Em=490/525nm.Readtheplateevery1–2secondsfor3minutes.
品牌介绍
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。
品牌分类
淋巴细胞信号传导 GPCR抗体 羰基活性生物素 细胞代谢 胺反应生物素 链霉亲和素结合物 生物素化抗IgGs 乙酰基特异性抗体 细胞骨架抗体
联络我们
服务热线:4000-520-616
(限工作日9:00-18:00)
QQ :1570468124
手机:18915418616