| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 492/514 |
| MW | N/A |
| CAS# | N/A |
| Solvent | Water |
| Storage | F/D/L |
| Category | CellAnalysis CellApoptosis |
| Related | ApoptosisandCytotoxicity BiochemicalAssays |
| Spectrum | AdvancedSpectrumViewer |
1. Culturecellstoadensityoptimalforapoptosisinductionaccordingtoyourspecificinductionprotocol,butnottoexceed2x106cells/mL.Atthesametime,cultureanon-inducednegativecontrolcellpopulationatthesamedensityastheinducedpopulationforeverylabelingcondition. Hereareafewexamplesfor inducingapoptosisinsUSPensionculture:
1)TreatingJurkatcellswith2μg/mlcamptothecinfor3hours.
2)TreatingJurkatcellswith1μMstaurosporinefor3hours.
3)TreatingHL-60cellswith4μg/mlcamptothecinfor4hours.
4)TreatingHL-60cellswith1μMstaurosporinefor4hours.
Note:Eachcelllineshouldbeevaluatedonanindividualbasistodeterminetheoptimalcelldensityforapoptosisinduction.
2. Make150XFAM-VDVAD-FMKDMSOstocksolutionbyadding50μLofDMSOtothevialofFAM-VDVAD-FMK(ComponentA).
3. Add150XFAM-VDVAD-FMKintothecellsolutionata1:150ratio,andincubatethecellsina37°C,5%CO2incubatorfor1hour.
Note1:Thecellscanbeconcentratedupto~5X106cells/mLforFAM-VDVAD-FMKlabeling.Theunused150XFAM-VDVAD-FMKDMSOstocksolutionshouldbedividedassingleusealiquotandstoredat-20°C.
Note2:Foradherentcells,gentlyliftthecellswith0.5mMEDTAtokeepthecellsintact,andwashthecellsoncewithserum-containingmediapriortoincubationwithFAM-VDVAD-FMK.
Note3:Theappropriateincubationtimedependsontheindividualcelltypeandcellconcentrationused.Optimizetheincubationtimeforeachexperiment.
4. Spindownthecellsat~200gfor5minutes,andwashcellswith1mLwashbuffer(ComponentB)twice.Resuspendthecellsindesiredamountofwashingbuffer.
Note1:FAM-VDVAD-FMKisfluorescent,thusitisimportanttowashoutanyunboundreagenttoeliminatethebackground.
Note2:Fordetachedcells,theconcentrationofcellsshouldbeadjustedto2-5X105cells/100μLaliquotpermicrotiterplatewellforuseinstep6.
5. Ifdesired,labelthecellswithaDNAstain(suchaspropidiumiodidefordeadcells,orHoechstforwholepopulationofthecellnucleusstain).
6. Monitorthefluorescenceintensitybyfluorescencemicroscopy,flowcytometer,orfluorescentmicroplatereaderatEx/Em=490/525nm(forpropidiumiodide,Ex/Em=535/635nm;forHoechstdyes,Ex/Em=350/461nm).
6.1 Forflowcytometry,monitorthefluorescenceintensityusingtheFL1channel(FL2channelforpropidiumiodidestaining).Gateonthecellsofinterest,excludingdebris.
6.2 Forfluorescencemicroscopyandfluorescentmicroplatereader.Place100μLofthecellsuspensionsintoeachofwellsofa96-wellblackmicrotiterplate.
Note: Ifitisnecessarytoequilibratethecellconcentrations,adjustthesuspensionvolumefortheinducedcellstoapproximatethecelldensityofthenon-inducedpopulation. Thisadjustmentstepisoptionalifyourcelltreatmentdoesnotresultinadramaticlossinstimulatedcellpopulationnumbers.
6.3 ObservecellsunderafluorescencemicroscopeusingFITCchannel(TRITCchannelforpropidiumiodidestaining,DAPIchannelforHoechststaining).
6.4 MonitorthefluorescenceintensityusingEx/Em=490/525nm(cutoffat515nm)bottomreadmodeusingafluorescentmicroplatereader.
| References&Citations |  CitationExplorer |
HelicobacterpyloriSecretedProteinHP1286TriggersApoptosisinMacrophagesviaTNF-IndependentandERKMAPK-DependentPathways
Authors:RaquelTavares,SushilKumarPathak
Journal:FrontiersinCellularandInfectionMicroBIOLOGy(2017):58
Deathreceptor3mediatesnecroptoticcelldeath
Authors:SebastianBittner,GertrudKnoll,MartinEhrenschwender
Journal:CellularandMolecularLifeSciences(2016):1--12
HelicobacterpyloriproteinJHP0290exhibitsproliferativeandanti-apoptoticeffectsingastricepithelialcells
Authors:RaquelTavares,SushilKumarPathak
Journal:PloSone(2015):e0124407
AAT Bioquest常用产品报价单
| 产品名 | 货号 | 公司分类 | 商城分类 | 美金价 |
| AAT Bioquest/Fluorescein, disodium salt *CAS 518-47-8*/2/100 mg | 2 | iFluor Rapid Tests | 酸碱缓冲液 | 1088.00 | AAT Bioquest/14-3-3 ζ (Ab-58) Antibody/8B0001/50 ug | 8B0001 | Signaling Intermediate Ab | 重组抗体 | 2828.00 | AAT Bioquest/Opioid Receptor (Phospho-Ser375) Antibody/8A0022/50 ug | 8A0022 | Phospho-Specific Ab | 受体 | 2828.00 | AAT Bioquest/iFluor™ 350 goat anti-mouse IgG (H+L)/16440/200 ug | 16440 | Fluorescent Anti-IgGs | 荧光染料 | 1088.00 | AAT Bioquest/trFluor™ Eu goat anti-mouse IgG (H+L)/16518/100 ug | 16518 | Fluorescent Anti-IgGs | 抗小鼠 | 4278.00 | AAT Bioquest/RPE-streptavidin conjugate/16900/100 ug | 16900 | Streptavidin Conjugates | 其他生物染料 | 1378.00 | AAT Bioquest/GCNT3 Antibody/8C14708/50 ug | 8C14708 | Signaling Intermediate Ab | 功能性抗体 | 2828.00 | AAT Bioquest/Amplite™ Luciferase Reporter Gene Assay Kit *Bright Glow*/12518/1 plate | 12518 | Reporter Gene Enzymes | 其它检测试剂盒 | 2103.00 | AAT Bioquest/ReadiLink™ Rapid mFluor™ Violet 450 Antibody Labeling Kit *Microscale Optimized for Labeling 50 &mi | 1100 | mFluor™ Dyes and Kits | 标记试剂盒 | 2103.00 | AAT Bioquest/Amplite™ Colorimetric Urea Quantitation Kit *Blue Color*/10058/200 Tests | 10058 | Diagnostic Molecules | 定量试剂盒 | 2828.00 | AAT Bioquest/Screen Quest™ CHO-Gqi Chimera Cell line/38101/Each | 38101 | cAMP GPCR Assays | 常规细胞株 | 0.00 |
本网站将在规定时间内给予删除等相关处理。
