>
产品中心 >
Other_kits >
AAT Bioquest/Cell Meter™ Caspase 3/7 Activity Apoptosis Assay Kit *Red Fluorescence*/22797/100 Tests
| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 534/619 |
| MW | N/A |
| CAS# | N/A |
| Solvent | N/A |
| Storage | F/D/L |
| Category | CellAnalysis CellCytotoxicity |
| Related | ApoptosisandCytotoxicity CellApoptosis BiochemicalAssays |
| Spectrum | AdvancedSpectrumViewer |
1.Preparecells:
1.1 Foradherentcells:Platecellsovernightingrowthmediumat20,000cells/well/90µLfora96-wellor5,000cells/well/20µLfora384-wellplateblackwall/clearbottomplate.
1.2 Fornon-adherentcells:CentrifugethecellsfromtheculturemediumandthensUSPendthecellpelletinculturemediumat80,000to200,000cells/well/90µLfora96-wellor20,000to50,000 cells/well/20µLfora384-wellblackwall/clearbottomplate.Centrifugetheplateat800rpmfor2minuteswithbrakeoffpriortotheexperiments.
Note:Eachcelllineshouldbeevaluatedonanindividualbasistodeterminetheoptimalcelldensityforapoptosisinduction.
2.Preparecaspase3/7assayloadingsolution:
2.1 Thawbothofthekitcomponentsatroomtemperaturebeforeuse.
2.2 Make200XZ-DEVD-ProRed™stocksolutionbyadding65µLDMSOintothevialofComponentA.
2.3 Makecaspase3/7assayloadingsolutionbyadding50µLofZ-DEVD-ProRed™Stocksolution(200X,fromStep
2.2) into10mLofAssayBuffer(ComponentB),andmixthemwell.
Note:AliquotandstoretheunusedZ-DEVD-ProRed™Stocksolution(fromStep2.2)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(3-4hoursforJurkatcellstreatedwithstaurosporine)toinduceapoptosis.
3.3 Add100µL/well/96-welor25µL/well/384-wellplateofcaspase3/7assayloadingsolution(fromstep2.3).
3.4 Incubatetheplateatroomtemperatureforatleast1hour,protectedfromlight.
Note:Ifdesired,add1µLofthe1mMAc-DEVD-CHOcaspase3/7inhibitortoselectedsamples10minutesbeforeaddingtheassayloadingsolutionatroomtemperaturetoconfirmtheinhibitionofthecaspase3/7-likeactivities.
3.5 MonitorthefluorescenceintensityatEx/Em=535/620nm(cutoffat610nm)witheithertoporbottomreadmode.
Note:Sometimes,bottomreadgivesbettersignaltobackgroundratio,centrifugecellplate(especiallyforthenon-adherentcells)at800rpmfor2minutes(brakeoff)ifusingbottomreadmode.
| References&Citations |  CitationExplorer |
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)
