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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
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当前位置: 首页 > 产品中心 > Other_kits > AAT Bioquest/Cell Meter™ Caspase 3/7 Activity Apoptosis Assay Kit *Green Fluorescence*/22796/200 Tests
商品详细AAT Bioquest/Cell Meter™ Caspase 3/7 Activity Apoptosis Assay Kit *Green Fluorescence*/22796/200 Tests
AAT Bioquest/Cell Meter™ Caspase 3/7 Activity Apoptosis Assay Kit *Green Fluorescence*/22796/200 Tests
AAT Bioquest/Cell Meter™ Caspase 3/7 Activity Apoptosis Assay Kit *Green Fluorescence*/22796/200 Tests
商品编号: 22796
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
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Ex/Em(nm)498/520
MWN/A
CAS#N/A
SolventN/A
StorageF/D/L
CategoryCellAnalysis
CellCytotoxicity
RelatedApoptosisandCytotoxicity
CellApoptosis
BiochemicalAssays
OurCellMeter™assaykitsareasetoftoolsformonitoringcellviABIlity.Thereareavarietyofparametersthatcanbeusedformonitoringcellviability.ThisparticularkitisdesignedtomonitorcellapoptosisthroughmeasuringCaspase3activation.Caspase3iswidelyacceptedasareliableindicatorforcellapoptosissincetheactivationofcaspase-3(CPP32/apopain)isimportantfortheinitiationofapoptosis.Caspase3hassubstrateselectivityforthepeptidesequenceAsp-Glu-Val-Asp(DEVD).ThiskitusesZ-DEVD-Rh110-DVED-Zasafluorogenicindicatorforcaspase-3activity.CleavageofRh110peptidesbycaspase3generatesstronglyfluorescentRh110thatismonitoredfluorimetricallyat520-530nmwithexcitationof480-500nm.Thekitprovidesalltheessentialcomponentswithanoptimizedassayprotocol.Theassayisrobust,andcanbereADIlyadaptedforhigh-throughputassays.Using100ulofreagentsperwellina96-wellformat,thiskitprovidessufficientreagentstoperform200assays.Using25ulofreagentsperwellina384-wellformat,thiskitprovidessufficientreagentstoperform800assays.
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Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.

1.Preparecells:

1.1   Foradherentcells:Platecellsovernightingrowthmediumat20,000cells/well/90µLfora96-wellplateor5,000cells/well/20µLfora384-wellplate.

 

1.2   Fornon-adherentcells:CentrifugethecellsfromtheculturemediumandthensUSPendthecellpelletinculturemediumat80,000cells/well/90µLfora96-wellpoly-Dlysineplateor20,000cells/well/20µLfora384-wellpoly-Dlysineplate.Centrifugetheplateat800rpmfor2minuteswithbrakeoffpriortotheexperiments.

Note:Eachcelllineshouldbeevaluatedonanindividualbasistodeterminetheoptimalcelldensityforapoptosisinduction.

 

2.Preparecaspase3/7assayloadingsolution:

2.1   Thawbothofthekitcomponentsatroomtemperaturebeforeuse.

 

2.2   Makecaspase3/7assayloadingsolutionbyadding50µLofCaspase3/7Substrate(ComponentA)into10mLofAssayBuffer(ComponentB),andmixwell.

Note:AliquotandstoretheunusedCaspase3/7Substrate(ComponentsA)andAssayBuffer(ComponentB)at-20oC.Avoidrepeatedfreeze/thawcycles.

 

3B3.Assayprocedures:

3.1   Treatcellsbyadding10µL/wellof10Xtestcompounds(96-wellplate)or5µL/wellof5Xtestcompounds(384-plate)intoPBSorthedesiredbuffer.Forblankwells(mediumwithoutthecells),addthesameamountofcompoundbuffer.

 

3.2   Incubatethecellplateina37°C,5%CO2,incubatorforadesiredperiodoftime(4-6hoursforJurkatcellstreatedwithcamptothecin)toinduceapoptosis.

 

3.3   Add100µL/well(96-wellplate)or25µL/well(384-wellplate)ofcaspase3/7assayloadingsolution(fromstep2.2).

 

3.4   Incubatetheassayloadingsolutionplateatroomtemperatureforatleast1hour,protectedfromlight.

Note:Ifdesired,add1µLofthe1mMAc-DEVD-CHOcaspase3/7inhibitortoselectedsamples10minutesbeforeaddingtheassayloadingsolutionatroomtemperaturetoconfirmtheinhibitionofthecaspase3/7-likeactivities.

 

3.5   Centrifugecellplate(especiallyforthenon-adherentcells)at800rpmfor2minutes(brakeoff).

 

3.6   MonitorthefluorescenceintensityatEx/Em=490/525nm.

References&Citations
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AngiopoietinsModulateSurvival,Migration,andtheComponentsoftheAng-Tie2PathwayofChronicLymphocyticLeukaemia(CLL)CellsInVitro
Authors:LuisMarioAguirrePalma,HannaFlamme,IrisGerke,Karl-AntonKreuzer
Journal:CancerMicroenvironment(2016):13--26

HypoxiaregulatesTRAILsensitivityofcolorectalcancercellsthroughmitochondrialautophagy.
Authors:GertrudKnoll,SebastianBittner,MariaKurz,JonathanJantsch,MartinEhrenschwender
Journal:Oncotarget(2016)

microRNA-186inhibitscellproliferationandinducesapoptosisinhumanesophagealsquamouscellcarcinomabytargetingSKP2
Authors:WeiHe,JianfangFeng,YanZhang,YuanyuanWang,WenqiaoZang,GuoqiangZhao
Journal:LaboratoryInvestigation(2016):317--324

Promotionofosteointegrationunderdiabeticconditionsbytantalumcoating-basedsurfacemodificationon3-dimensionalprintedporoustitaniumimplants
Authors:LinWang,XiaofanHu,XiangyuMa,ZhenshengMa,YangZhang,YizhaoLu,XiangLi,WeiLei,YafeiFeng
Journal:ColloidsandSurfacesB:Biointerfaces(2016):440--452

Roleofdelta-likeligand-4inchemoresistanceagainstdocetaxelinMCF-7cells
Authors:QWang,YShi,HJButler,JXue,GWang,PDuan,HZheng
Journal:Human&ExperimentalToxicology(2016):0960327116650006

Glucosepromotescellproliferation,glucoseuptakeandinvasioninendometrialcancercellsviaAMPK/mTOR/S6andMAPKsignaling
Authors:JianjunHan,LuZhang,HuiGuo,WeiyaZWysham,DarioRRoque,AdamKWillson,XiuguiSheng,ChunxiaoZhou,VictoriaLBae-Jump
Journal:Gynecologiconcology(2015):668--675

IL-7abrogatestheimmunosuppressivefunctionofhumandouble-negativeTcellsbyactivatingAkt/mTORsignaling
Authors:AndreaAllgäuer,ELISAbethSchreiner,FulviaFerrazzi,ArifBEkici,ArminGerbitz,AndreasMackensen,SimonVölkl
Journal:TheJournalofImmunology(2015):3139--3148

InsulinimprovesosteogenesisoftitaniumimplantsunderdiabeticconditionsbyinhibitingreactiveoxygenspeciesoverproductionviathePI3K-Aktpathway
Authors:LinWang,XiongZhao,Bo-yuanWei,YiLiu,Xiang-yuMa,JianWang,Peng-chongCao,YangZhang,Ya-boYan,WeiLei
Journal:Biochimie(2015):85--93

LGL1modulatesproliferation,apoptosis,andmigrationofhumanfetallungfibroblasts
Authors:HuiZhang,NeilBSweezey,FeigeKaplan
Journal:AmericanJournalofPhysiology-LungCellularandMolecularPhysiology(2015):L391--L402

t-BHQProvidesProtectionagainstLeadNeurotoxicityviaNrf2/HO-1Pathway
Authors:FangYe,XiaoyiLi,LiliLi,JingYuan,JunChen
Journal:Oxidativemedicineandcellularlongevity(2015)


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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
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