SampleTyple | Plasma,Serum,Cellculturesupernatant,otherbodyfluid | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Contents | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Method | Colorimetricmethodat490nm | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
StandardCurve | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sensitivity | 8µM-400µM | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ReactionTime | 15minutes | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Applications | ForBIOLOGicalresearch: | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Storage | -80°C | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Notes | Materialsneededbutnotsupplied Aplatereadercapableofmeasuringabsorbanceat490nm AdjustablePipettesandarepeatpipettor Distilledwater(milliQorHPLC-grade) 0.5MAceticAcid Clearflatbottom96-wellplatesifnotincludedinthekitpurchased | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Shipping | Icepacks |
ReagentPreparation Note:Allreagentsarefrozen.Werecommendyouspinthestandardvialbeforeopening. 1.GlucoseStandard(2x) Note:Pleasedon’tdodilutionintheprovidedGlucosestandardvialitself. Thevialcontains500µlof800µMGlucoseStandard.Thestandardmustbeequilibratedtoroomtemperaturebeforeuse.Dilute500µlof800µMGlucoseStandardwith500µlofdH2Otopreparea400μMGlucoseStandard.1mlofdilutedstandardisenoughformaking3standardcurvesifassayedinduplicate.Storeat-80?C. 2.GlucoseAssaySolution Thesolutioncontainsenzymesthatarelightsensitive.Thesolutionmustbethawedonicebeforeuse.Besttoaliquottheamountneededanduseitalltopreventthawing/freezingcycles.Freezeandstorealiquotsat-80oC | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Protocol 1.SamplePreparation Serum/Plasma/otherbodyfluid/cellculturesupernatant Note: | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
2.StandardCurvePrepation WerecommendthatGlucoseStandardsbeassayedinduplicate.Astandardcurvehastoberunineachassay. | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
3.Performtheassay | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
4.Calculation a.AveragetheOD490nmvaluesofreplicatewellsofeachGlucosestandard,testsamples,andblank.Inordertogetthecorrectedabsorbance,subtracttheaverageOD490nmvalueoftheblankfromtheaveragedOD490nmvaluesfromallstandardsandsamples. Glucose(μM)=[(Correctedabsorbance)-(y-intercept)]/Slope MaterialSafetyDataSheet Date:Oct26,2015 1.PRODUCTANDCOMPANYIDENTIFICATION 1.1Productidentifier Productname:GlucoseAssayKitI CatalogNo:1200031002,120003100A,120003100P,1200032002,120003200A,12003200P,1200034002,120003400A,120003400P 1.2Relevantidentifiedusesofthesubstanceormixtureandusesadvisedagainst Forresearchuseonly. 1.3Detailsofthesupplierofthesafetydatasheet Company:EtonBioscienceInc TollFree:1-800-758-1630 Tel:1-800-758-1630 Fax:1-800-507-2912 1.4Emergencytelephonenumber 1-800-758-1630 2.HAZARDSIDENTIFICATION 2.1Classificationofthesubstanceormixture Notahazardoussubstanceormixture 2.2GHSLabelelements,includingprecautionarystatements Notahazardoussubstanceormixture 2.3Otherhazards None 3.COMPOSITION/INFORMATIONONINGREDIENTS 3.1Substances
9.PHYSICALANDCHEMICALPROPERTIES 9.1Informationonbasicphysicalandchemicalproperties
9.2Othersafetyinformation Nodataavailable. 10.STABILITYANDREACTIVITY 10.1Reactivity Nodataavailable 10.2Chemicalstability Stableunderrecommendedstorageconditions. 10.3Possibilityofhazardousreactions Nodataavailable. 10.4Conditionstoavoid Nodataavailable 10.5IncompatIBLematerials Strongoxidizingagents 10.6Hazardousdecompositionproducts Hazardousdecompositionproductsformedunderfireconditions:carbonoxides 11.TOXICOLOGICALINFORMATION 11.1Informationontoxicologicaleffects Toxicologicaleffectsonthisproducthavenotbeenthoroughlystudied. 12.ECOLOGICALINFORMATION 12.1Toxicity Avoidreleaseintoenvironment 12.2Persistenceanddegradability Nodataavailable 12.3.Bioaccumulativepotential Nodataavailable 12.4Mobilityinsoil Nodataavailable 12.5ResultsofPBTandvPvBassessment Nodataavailable 12.6Otheradverseeffects Nodataavailable 13.DISPOSALINFORMATION 13.1Wastetreatmentmethods Disposeinaccordancewithlocal,state,andfederalregulations. 14.TRANSPORTINFORMATION DOT: Propershippingname:none Propershippingname:none ADR/RID Propershippingname:none 15.REGULATORYINFORMATION EURiskandSafetyphrases: R36/37/38:irritatingtoeyes/respiratorysystem/skin 16.OTHERINFORMATION Theinformationaboveisbelievedtobeaccurateandrepresentsthebestinformationcurrentlyavailabletous.However,wemakenowarrantyofmerchantabilityoranyotherwarranty,expressorimplied,withrespecttosuchinformation.EtonBioscienceInc.shallnotbeheldliableforanydamagesorotherconsequencesresultingfromhandlingorfromcontactwiththeaboveproduct.ZakikhaniM,BazileM,HashemiS,JaveshghaniS,AvizonisD,etal.(2012)AlterationsinCellularEnergyMetabolismAssociatedwiththeAntiproliferativeEffectsoftheATMInhibitorKU-55933andwithMetformin.PLoSONE7(11):e49513.doi:10.1371/journal.pone.0049513 ShuangLiang,*PaolaGalluzzo,*AnnaSobol,SylviaSkucha,BrittanyRambo,andMaurizioBocchetta.MultimodalityApproachestoTreatHypoxicNon–SmallCellLungCancer(NSCLC)Microenvironment.GenesCancer.2012February;3(2):141–151. ZHANG,YANBINetal.GlucoseTransporter3PerformsaCriticalRoleinmTOR-MediatedOncogenicGlycolysisandTumOrigenesis.OncologyLetters9.6(2015):2809–2814.PMC.Web.6Oct.2015. EyalAmiel,BartEverts,ToriC.Freitas,IrahL.King,JonathanD.Curtis,ErikaL.PearceandEdwardJ.Pearce.InhibitionofMechaNISTicTargetofRapamycinPromotesDendriticCellActivationandEnhancesTherapeuticAutologousVaccinationinMice.JImmunol2012;189:2151-2158 ZhiguoLi,JieLi,PenpengBi,YingLu,GrantBurcham,BennetD.Elzey,TimothyRatliff,StephenF.Konieczny,NihalAhmad,ShihuanKuang,andXiaoquiLiu.Plk1PhosphorylationofPTENcausesatumor-PromotingMetabolicState.Mol.Cell.Biol.October2014vol.34no.193642-3661 QingsongCai,TongLin,SushamaKamarajugadda,andJianrongLu.RegulationofGlycolysisandtheWarburgEffectbyEstrogen-relatedReceptors.Oncogene.2013April18;32(16):2079–2086. WhattypeofmediumshouldIuseformakingculturedcellsforthisassay? Pleaseusephenolredfreemedium.Pleasedonotusephenolredmediumsincephenolredwouldaffectabsorbancereadings. Whatenzymeisusedinthisassay? ItisGlucoseOxidase. Whatarethecompositionsofthisassay? Unfortunatelyitisproprietary. Whatisthesensitivityofthisassay? Itis8μM-400μM. DoIneedtomakeastandardcurveeverytime? Yesitisnecessarysinceyoucalculateyoursampleconcentrationbasedonthestandardcurve.Pleasedonotusetherepresentativestandardcurveintheprotocoltocalculateconcentrationsofyoursamples. HowmuchsampledoIneedforthisassay? End-userswouldhavetoperformanexperimenttodetermineoptimalvolumeofsampleswhosereadingsfallwithinthestandardcurverange.Pleaserefertopublicationswithwhichthisproductwasusedtoseehowmuchsampleshavebeenusedinsimilarexperiments. HowcanIdilutemysamples? YoursamplesmaybedilutedwithPBSordH2O. CanIusefluorescencespectroscopytomeasurereadingsforthisassay? No,thekitdoesnotworkwithafluorescencereadersincethekitemployscolorimetricassays. CanIusefrozensamples? Althoughitisbettertousefreshsamplesforassays,youcanusefrozensamples.Ifyouarenotusingfrozensamplesinonce,pleasemakealiquotsofyoursamplesbeforeyouputtheminafreezertopreventfromdegradingsamplesfromrepeatedfreeze-thawcycles. Mysampleformedprecipitationafteraddingaceticacid.HowcanIpreventthis? Youcanstillmeasurereadingswithoutaddingaceticacid. Theprotocolrecommendsmeasuringtheabsorbanceat490nm.Is485nmcloseenough? Itisokaslongasyousetuptheabsorbancebetween470and490nm. Whyaremystandardcurvevaluesdifferentthantheoneshownonthedatasheet? Manyfactorsinfluencethevaluessuchasroomtemperature,incubationtime,handling,etc. Aretrialkitsavailable? Yestheyareavailableforfirsttimecustomers;however,youwillberesponsibleforshippingandhandlingfee.Pleasecontact formoredetails.
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