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/Amplite™ Fluorimetric Proteasome 20S Activity Assay Kit *Green Fluorescence*/13456/100 tests
产品编号:13456
市  场 价:¥85560.00
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:$4278.00
品      牌: AAT Bioquest
公      司:AAT Bioquest
公司分类:
AAT Bioquest/Amplite™ Fluorimetric Proteasome 20S Activity Assay Kit *Green Fluorescence*/13456/100 tests
商品介绍
Overview
PrinterFriendlyVersion

Ex/Em(nm)498/520
MWN/A
CAS#N/A
SolventN/A
StorageF/D/L
CategoryEnzymeDetection
PeptidasesandProteases
RelatedBiochemicalAssays
OurAmplite™Fluorometric20SProteasomeAssayKitisahomogeneousfluorescentassaythatmeasuresthechymotrypsin-likeproteaseactivityassociatedwiththeproteasomecomplexeitherinculturedcellsorcelllysates.ThiskitusesLLVY-R110asafluorogenicindicatorforproteasomeactivity.CleavageofLLVY-R110byproteasomegeneratesthestronglygreenfluorescentR110thatismonitoredfluorimetricallyat520-530nmwithexcitationof480-500nm.Thekitprovidesalltheessentialcomponentswithanoptimizedassayprotocol.Theassayisrobust,andcanbereADIlyadaptedforhigh-throughputassaysforevaluationofproteasomeactivityorinhibitorscreeninginculturedcellsorinsolution.Theassaycanbeperformedinaconvenient96-welland384-wellfluorescencemicrotiter-plateformat.Themainfunctionoftheproteasomeistodegradeunneededordamagedproteinsbyproteolysis,achemicalreactionthatbreakspeptidebonds.Theproteasomaldegradationpathwayisessentialformanycellularprocesses,includingthecellcycle,theregulationofgeneexpression,andresponsestooxidativestress.Themostcommonformoftheproteasomeinthispathwayisthe26Sproteasome,anATP-dependentproteolyticcomplex,whichcontainsone20S(700-kDa)coreparticlestructureandtwo19S(700-kDa)regulatorycaps.The20Scorecontainsthreemajorproteolyticactivitiesincludingchymotrypsin-like,trypsin-likeandcaspase-like.ItisresponsIBLeforthebreakdownofkeyproteinsinvolvedwithapoptosis,DNArepair,endocytosis,andcellcyclecontrol.
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:

 

1.1   Foradherentcells:Platecellsovernightingrowthmediumat80,000cells/well/90µLfora96-wellplateor20,000cells/well/20µLfora384-wellplate.

 

1.2   Fornon-adherentcells:CentrifugethecellsfromtheculturemediumandthensUSPendthecellpelletinculturemediumat300,000cells/well/90µLfora96-wellpoly-Dlysineplateor80,000cells/well/20µLfora384-wellpoly-Dlysineplate.Centrifugetheplateat800rpmfor2minuteswithbrakeoffpriortotheexperiments.

Note:Eachcelllineshouldbeevaluatedonanindividualbasistodeterminetheoptimalcelldensity.

 

2.Prepareproteasomeassayloadingsolution:

 

2.1   Thawallthekitcomponentsatroomtemperaturebeforeuse.

 

2.2   Make400XProteasomeLLVY-R110Substratestocksolution:Add25µLofDMSO(ComponentC)tothevialofProteasomeLLVY-R110Substrate(ComponentA),andmixwell.

 

2.3   Makeproteasomeassayloadingsolution:Add25µLof400XProteasomeLLVY-R110Substratestocksolution(fromStep2.2)into10mLofAssayBuffer(ComponentB),andmixwell.

Note:25µLof400XProteasomeLLVY-R110Substratestocksolution(fromStep2.2)and10mLofAssayBuffer(ComponentB)areenoughfor1plate.Aliquotandstoretheunused400XProteasomeLLVY-R110SubstratestocksolutionandAssayBufferat-20oC.Avoidrepeatedfreeze-thawcycles.

 

3.Runtheproteasomeassay:

 

3.1   Treatcellswith10µLof10Xtestcompound(fora96-wellplate)or5µLof5Xtestcompound(fora384-wellplate)inPBSordesiredbuffer.Forblankwells(mediumwithoutthecells),addthecorrespondingamountofcompoundbuffer.

 

3.2   Incubatethecellplatesina5%CO2,37°Cincubatorforadesiredperiodoftime.

Note:Pureproteasomeorcelllysatescanbeuseddirectlyforscreeningtheproteasomeinhibitors.

 

3.3   Add100µL/well(96-wellplate)or25µL/well(384-wellplate)ofproteasomeassayloadingsolution(fromStep2.3).

 

3.4   Incubatetheplateat37°Corroomtemperatureforatleast1hour(2hourstoovernight),protectedfromlight.

Note:Eachcelllineshouldbeevaluatedonanindividualbasistodeterminetheoptimalincubationtime.

 

3.5   Monitorthefluorescenceintensity(topread)atEx/Em=490/525nm.

References&Citations
CitationExplorer

MycobacteriumtuberculosisControlsPhagosomalAcidificationbyTargetingCISH-MediatedSignaling
Authors:ChristopheJQueval,Ok-RyulSong,Jean-PhilippeCarralot,Jean-MichelSaliou,AntoninoBongiovanni,GaspardDeloison,NathalieDeboosère,SamuelJouny,RaffaellaIantomasi,VincentDelorme
Journal:CellReports(2017):3188--3198

Autophagy-mediatedclearanceofubiquitinatedmutanthuntingtinbygrapheneoxide
Authors:PeipeiJin,PengfeiWei,YunjiaoZhang,JunLin,RuiSha,YiHu,JiqianZhang,WeiZhou,HanYao,LiRen
Journal:Nanoscale(2016)

InteractomeanalysisrevealsanovelroleforRAD6intheregulationofproteasomeactivityandlocalizationinresponsetoDNAdamage
Authors:HongliAn,LuYang,ChenWang,ZhixueGan,HaihuiGu,TaoZhang,XinHuang,YanLiu,YufengLi,Shing-JyhChang
Journal:MolecularandCellularBIOLOGy(2016):MCB--00419

L-BMAAinducedERstressandenhancedcaspase12cleavageinhumanneuroblastomaSH-SY5Ycellsatlownonexcitotoxicconcentrations
Authors:OliverOkle,KerstinStemmer,UlrichDeschl,DanielRDietrich
Journal:toxicologicalsciences(2013):217--224


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