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 |
| |||||||||||||||||||||
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.
ProductCitations(0)
Currentlytherearenocitationsforthisproduct.