请使用支持JavaScript的浏览器!
主营:研究并生产荧光和发光探针,信号转导研究的试剂
℡ 4000-520-616
℡ 4000-520-616
AAT Bioquest/Cell Meter™ Fluorimetric Intracellular Peroxynitrite Assay Kit *Green Fluorescence*/16315/100 Tests
产品编号:16315
市  场 价:¥100060.00
场      地:美国(厂家直采)
联系QQ:1570468124
电话号码:4000-520-616
邮      箱: info@ebiomall.com
美  元  价:$5003.00
品      牌: AAT Bioquest
公      司:AAT Bioquest
公司分类:
AAT Bioquest/Cell Meter™ Fluorimetric Intracellular Peroxynitrite Assay Kit *Green Fluorescence*/16315/100 Tests
商品介绍
Overview
PrinterFriendlyVersion

Ex/Em(nm)502/535
MWN/A
CAS#N/A
SolventDMSO
StorageF/D/L
CategoryNeuroBIOLOGy
ReactiveOxygenSpecies
RelatedCellSignaling
BiochemicalAssays
Peroxynitrite(ONOO-)isastrongoxidizingspeciesandahighlyactivenitratingagent.PeroxynitriteisformedfromthereactionbetweensuperoxiderADIcalsandnitricoxidegeneratedincells.Itcancausedamagestoawidearrayofbiomoleculesincludingproteins,enzymes,lipidsandnucleicacids,eventuallycontributingtocelldeath.Meanwhile,peroxynitritecanalsohaveprotectiveactivitiesinvivobycontributingtohost-defenseresponsesagainstinvadingpathogens.Therefore,peroxynitriteisanessentialbiologicaloxidantinvolvedinaboardrangeofphysiologicalandpathologicalprocesses.Duetoitsextremelyshorthalf-lifeandlowsteady-stateconcentration,ithasbeenchallengingtodetectandunderstandtheroleofperoxynitriteinbiologicalsystems.AATBioquest"sDAX-J2™PONGreenhasbeendevelopedtoaddressthisumetneed.ItprovidesasensitivetooltomonitorONOO-levelinlivingcells.AATBioquest"sDAX-J2™PONGreenspecificallyreactswithintercellularONOO-togenerateabrightgreenfluorescentproduct.Itcanbeusedinfluorescenceimaging,flowcytometryandfluorescencemicroplatereader-basedassays.
SpectrumAdvancedSpectrumViewer



Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.

1.Preparecells:

1.1.    Foradherentcells:Platecellsovernightingrowthmediumat20,000to80,000cells/well/90μLfora96-wellplateor5,000to20,000cells/well/22.5μLfora384-wellplate.

1.2.    Fornon-adherentcells:CentrifugethecellsfromtheculturemediumandsUSPendthecellpelletsinculturemediumat80,000-200,000cells/well/90µLfora96-wellpoly-Dlysineplateor20,000-50,000cells/well/22.5µLfora384-wellpoly-Dlysineplate.Centrifugetheplateat800rpmfor2minuteswithbrakeoffpriortoyourexperiment.

Note:Eachcelllineshouldbeevaluatedonanindividualbasistodeterminetheoptimalcelldensity.

2.Prepareworkingsolution:

2.1   PrepareDAX-J2™PONGreenstocksolution(500X):Add20µLofDMSO(ComponentC)intothevialofDAX-J2™PONGreen(ComponentA),andmixthemwell. 

Note:20μLofreconstitutedDAX-J2™PONGreenstocksolutionisenoughfor1plate.Unusedportioncanbealiquotedandstoredat≤-20°Cformorethanonemonthifthetubesaresealedtightlyandkeptfromlight.Avoidrepeatedfreeze-thawcycles.

2.2   PrepareDAX-J2™PONGreenworkingsolution(10X):Add10μLof500XDMSOreconstitutedDAX-J2™PeroxynitriteSensorstocksolution(fromStep2.1)into500μLofAssayBuffer(ComponentB),andmixthemwell.

Note:Theworkingsolutionisnotstable;prepareitasneededbeforeuse.

 

 

3.Runtheperoxynitriteassay:

3.1   Add10µL/well(96-wellplate),or2.5µL/well(384-wellplate)ofDAX-J2™PONGreenworkingsolution(fromStep2.2)in90µL(96-wellplate)or22.5µL(384-wellplate)cellcultureperwellinthecellplate(fromStep1).

 Note:Itisnotnecessarytowashcellsbeforestaining.It’srecommendedtostainthecellsinfullmedium.

3.2   Co-incubatecellswithDAX-J2™PONGreenwithtestcompoundsinfullmediumorinyourdesiredbufferat37ºCfordesiredperiodoftime,protectedfromlight.Forcontrolwells(untreatedcells),addthecorrespondingamountofcompoundbuffer.

Note1:It’srecommendedtostainthecellsinfullmedium.However,iftestedcompoundsareserumsensitive,growthmediumandserumfactorscanbeaspiratedawaybeforestaining.Add90µL/well(96-wellplate)and22.5µL/well(384-wellplate)of1XHank’ssaltsolutionand20mMHepesbuffer(HHBS)orthebufferofyourchoiceafteraspiration.Alternatively,cellscanbestainedinserum-freemedia.

Note2:Weco-incubatedRAW264.7macrophagecellswith50-200μMSIN-1andDAX-J2™PONGreeninfullmediumat37ºCfor1hourtoinduceperoxynitrite.SeeFigure1fordetails.

3.3   Alternatively,staincellswithDAX-J2™PONGreenat37ºCfor1hour,protectedfromlight(asinStep3.1).Removetheworkingsolution,thentreatcellswithtestcompoundsinfullmediumorinyourdesiredbufferat37ºCfordesiredperiodoftime.

3.4   MonitorthefluorescenceincreaseusingmicroplatereaderatEx/Em=490/530nm(cutoff=515nm)withbottomreadmode,ortakeimagesusingfluorescencemicroscopewithaFITCfilter.

References&Citations
CitationExplorer

Fluorescentreal-timequantitativemeasurementsofintracellularperoxynitritegenerationandinhibition
Authors:ZhenLuo,QinZhao,JixiangLiu,JinfangLiao,RuoguPeng,YuntingXi,ZhenjunDiwu
Journal:Analyticalbiochemistry(2017):44--48

GenesilencingofendothelialvonWillebrandFactorattenuatesangiotensinII-inducedendothelin-1expressioninporcineaorticendothelialcells
Authors:AnarDushpanova,SilviaAgostini,EnricaCiofini,ManuelaCABIati,ValentinaCasieri,MarcoMatteucci,SilviaDelRy,AldoClerico,SergioBerti,VincenzoLionetti
Journal:ScientificReports(2016)


品牌介绍

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 mg2iFluor Rapid Tests酸碱缓冲液1088.00
AAT Bioquest/14-3-3 ζ (Ab-58) Antibody/8B0001/50 ug8B0001Signaling Intermediate Ab重组抗体2828.00
AAT Bioquest/Opioid Receptor (Phospho-Ser375) Antibody/8A0022/50 ug8A0022Phospho-Specific Ab受体2828.00
AAT Bioquest/iFluor™ 350 goat anti-mouse IgG (H+L)/16440/200 ug16440Fluorescent Anti-IgGs荧光染料1088.00
AAT Bioquest/trFluor™ Eu goat anti-mouse IgG (H+L)/16518/100 ug16518Fluorescent Anti-IgGs抗小鼠4278.00
AAT Bioquest/RPE-streptavidin conjugate/16900/100 ug16900Streptavidin Conjugates其他生物染料1378.00
AAT Bioquest/GCNT3 Antibody/8C14708/50 ug8C14708Signaling Intermediate Ab功能性抗体2828.00
AAT Bioquest/Amplite™ Luciferase Reporter Gene Assay Kit *Bright Glow*/12518/1 plate12518Reporter 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 &mi1100mFluor™ Dyes and Kits标记试剂盒2103.00
AAT Bioquest/Amplite™ Colorimetric Urea Quantitation Kit *Blue Color*/10058/200 Tests10058Diagnostic Molecules定量试剂盒2828.00
AAT Bioquest/Screen Quest™ CHO-Gqi Chimera Cell line/38101/Each38101cAMP GPCR Assays常规细胞株0.00
自营商城图标
厂家直采
全球直采 正品优价
正品保障图标
正品保障
厂家直发 有线跟踪
解放采购图标
正规清关
CIF100%正规报关,提供发票
及时交付图标
及时交付
限时必达 不达必赔