
Overview:
ProductName | 20SProteasomeActivityKit |
Description | Fluorometricdetectionofpurified20Sproteasomeactivity |
SpeciesReactivity | SpeciesIndependent |
Platform | Microplate |
SampleTypes | PurifiedProteins |
DetectionMethod | FluorometricAssay |
AssayType | ContinuousKineticAssay |
Utility | Activitykitusedtoscreenpotentialproteasomeinhibitorsandactivators,andtomeasureandquantify20Sproteasomeactivityincontinuouskineticandend-pointassays. |
IncubationTime | 15-30minutes |
NumberofSamples | 40samplesinduplicateusingtheplateformat,or100x50µLmicro-cuvettebasedassays |
OtherResources | KitBooklet,MSDS |
Properties
StorageTemperature | 4ºC | |||||||||||||||||||||
ShippingTemperature | BlueIce | |||||||||||||||||||||
ProductType | ActivityKits | |||||||||||||||||||||
AssayOverview | The20SProteasomeActivitykitfacilitatestherapid,robustmeasurementofproteasomeactivityinvitro.Thekitutiliseshighpurity,fluorogenicsubstrateSuc-LLVY-AMCtogetherwithsuitablecalibrationstandardsandcontrolsfortheaccurateandsensitiveassessmentofproteasomechymotrypsin-like(β5)activity.Continuouskineticorend-pointassayscanbeperformedinmicro-cuvettesorin96-wellplateformatformulti-sampleanalysis.Containssufficientmaterialsforonefull96-wellplateassayorupto100x50µLmicro-cuvettebasedassaystoberun. | |||||||||||||||||||||
KitOverview |
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CiteThisProduct | 20SProteasomeActivityKit(StressMarqBiosciencesInc.,VictoriaBCCANADA,Catalog#SKT-133) |
BIOLOGicalDescription
AlternativeNames | Chymotrypsin-like(β5)ActivityKit |
ResearchAreas | Cancer,Apoptosis,CardiovascularSystem,CellSignaling,Neurodegeneration,Neuroscience,Post-translationalModifications |
ScientificBackground | Proteasomesarenon-lysosomalproteolyticcomplexeslocalisedprimarilyinthecytoplasmandinthenucleusofeukaryoticcells.TheyareresponsIBLefortheubiquitin-mediateddegradationofshorthalf-lifeproteinsandpeptidesthatareinvolvedinessentialcellularprocessesincludingcell-cycleregulation,apoptosisandtranscriptionalregulation,innateimmunityandantigenprocessing,andintheremovalofredundantordamagedproteins.Assuchproteindegradationbytheubiquitin-proteasomepathwayhasamajorregulatoryfunctionforproliferationactivityandsurvivalofbothnormalandmalignantcells,anditsdysfunctionhasbeenimplicatedinawiderangeofotherdiseaseprocessesincludingneurodegenerative,cardiovascularandmetabolicdisorders.The26Sproteasomestructureiscomposedofa20Sproteasomecatalyticcorecomplexandoneortwo19S(PA700)regulatorysubcomplexes.The20Scorecomprisestwocopiesof14subunits(7alphasubunitsand7betasubunits)arrangedinaα7β7β7α7cylindricalarray.Proteolyticactivitiesaredeterminedbytheβ1(caspase-like),β2(trypsin-like)andβ5(chymotrypsin-like)subunits,accesstowhichisguardedbytheα-subunits.The19SregulatoryunitconsistsofsixATPaseandatleasttennon-ATPasesubunitsthatarerequiredforubiquitinatedproteinbinding,deubiquitination,substrateunfoldingandtranslocationtothe20Scatalyticcore.Varyingcatalyticsubunitcomposition(β1,β1i;β2,β2i;β5,β5i)resultsinavarietyofpossiblesubtypesfromfullconstitutiveproteasome(β1,β2,β5)throughmixedpopulationstofullinducible/immunoproteasome(β1i,β2i,β5i).AlternativeregulatorycomplexessuchasthePA200and11Sproteasomeactivatorsconferdifferentsubstratespecificitiesandactivitycomparedtothe19Sregulator. |
References | 1.Saeki,Y.&Tanaka,K.Methodsinmolecularbiology(Clifton,N.J.)832,315–37(2012). 2.Frankland-Searby,S.&Bhaumik,S.R.Biochimicaetbiophysicaacta1825,64–76(2012). 3.Basler,M.,Kirk,C.J.&Groettrup,M.Currentopinioninimmunology1–7(2012). 4.Löw,P.Generalandcomparativeendocrinology172,39–43(2011). 5.Dennissen,F.J.a,Kholod,N.&VanLeeuwen,F.W.Progressinneurobiology96,190–207(2012). 6.Geng,F.,Wenzel,S.&Tansey,W.P.Annualreviewofbiochemistry81,177–201(2012). |
ProductImages

AMCStandardCurvefor20SProteasomeActivityKit.Representativeplotoffluorescencemeasurements(RFU)ofserialdilutionsofAMCStandardatrecommendedconcentrations.

Continuouskineticenzymeactivityassayshowingthetimecourseofcontrol20SproteasomeactivitywithSuc-LLVY-AMCsubstrateusingtheStressXpress®20SProteasomeActivityKit.20Sproteasome(~2.5nM)wasincubatedwith100µMSuc-LLVY-AMCinproteasomeassaybufferfor30minutesatroomtemperaturealongsideasubstrateonly(blank)controlreaction.Fluorescencemeasurements(RFU)weretakenat30secondintervalsandplottedvs.time.
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我使用的是1%琼脂糖凝胶,电泳缓冲液为0.5×TBE。
请问各位大虾,在不影响实验结果的前提下,这样的含EB的电泳缓冲液可以反复使用多少次?
谢谢^_^
此外,我们常常用“电泳法”判断液体的性质,是胶体还是溶液。在高中化学中我们就用过这种方法判断给定的液体是否为胶体。MOPS电泳缓冲液、胚胎干细胞培养生长因子、动物肝细胞分离培养试剂盒、结晶紫细胞群落染色试剂盒、MFN-2蛋白表达检测试剂盒、血红细胞溶解液、磷酸缓冲盐溶液、Hanks平衡盐粉剂、胰蛋白酶溶液、
此外,我们常常用“电泳法”判断液体的性质,是胶体还是溶液。在高中化学中我们就用过这种方法判断给定的液体是否为胶体。向左转|向右转

