SampleTyple | Plasma,Serum,Cellculturesupernatant,otherbodyfluid | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Contents | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Method | Colorimetricmethodat490nm | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
StandardCurve | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sensitivity | 60µM-3000µM | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ReactionTime | 30minutes | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Applications | ForBIOLOGicalresearch: | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Storage | -80°C | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Notes | Materialsneededbutnotsupplied Aplatereadercapableofmeasuringabsorbancebetween470-490nm AdjustablePipettesandarepeatpipettor Asourceofpurewater;glassdistilledwaterofHPLC-gradewaterissufficient 0.5MAceticAcid Clearflatbottom96-wellplatesifnotincludedinthekitpurchased | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Shipping | Icepacks | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ReagentPreparation Note:Allreagentsarefrozen.SWerecommendyouspinsmallvialsbeforeopening.
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Protocol 1.SamplePreparation Serum/Plasma/otherbodyfluid/cellculturesupernatant Note: | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
2.StandardCurvePreparation | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
3.Performtheassay a)Add50ulofL-LactateassaysolutiontoeachwellcontainingL-Lactatestandardsandtestsamples. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
4.Calculations a)AveragetheOD490nmvaluesofreplicatewellsofeachL-Lactatestandard,testsamples,andblank.Inordertogetthecorrectedabsorbance,subtracttheaverageOD490nmvalueoftheblank(L-LactateStandard#8)fromtheaverageOD490nmvaluesfromallstandardsandsamples. b)MakeastandardcurvebyplottingOD490nmvaluesfromeachL-LactatestandardsasafunctionofL-Lactateconcentration.Thiscanbedonewithexcelspreadsheet.CalculatethevalueofL-Lactateinsamplesusingtheequationobtainedfromthelinearregressionofthestandardcurve. L-Lactate(µM)=[(Correctedabsorbance)-(y-intercept)]/slope MaterialSafetyDataSheet Date:Oct26,2015 1.PRODUCTANDCOMPANYIDENTIFICATION 1.1Productidentifier Productname:L-LactateAssayKitI CatalogNo:1200011002,120001100A,120001100P,1200012002,120001200A,12001200P,1200014002,120001400A,120001400P 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 L-LactateAssaySolution
L-LactateStandard
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.Q.WhattypeofmediumshouldIuseformakingculturedcellsforLactateassay? A.Pleaseusephenolredfreemedium.Pleasedonotusephenolredmediumsincephenolredwouldaffectabsorbancereadings. Q.WhatenzymesisusedinLactateAssay? A.ItisLDH. Q.CanyoutellmethecompositionsofyourLactateAssay? A.Unfortunatelyitisproprietary. Q.HowdoIknowwhetherIneedtouseL-LactateorD-Lactateassaykit? A.L-Lactateassaykitsarecommonlyusedformeasuringlactatelevelinanimalcells. D-lactateassaykitsarecommonlyusedformeasuringlactatelevelinbacteriacells. Q.DoIneedtomakeastandardcurveeverytime? A.Yes,itisnecessarysinceyoucalculateyoursampleconcentrationbasedonthestandardcurve.PleasedonotusetherepresentativeLactatestandardcurveintheprotocoltocalculateconcentrationsofyoursamples. Q.CanIusefluorescencespectroscopytomeasurereadingsforLactateassaykits? A.No,thekitdoesnotworkwithafluorescencereadersincethekitemployscolorimetricassays. Q.HowdoImeasureLactatefromculturedcells? A.Youcangrowcellsinphenolred-freemedium,thenyoucanextractcellswith80%ethanol. Q.CanIusefrozensamples? A.Althoughitisbettertousefreshsamplesforassays,youcanusefrozensamples.Ifyouarenotusingfrozensamplesinonce,pleasemakealiquotsofyoursamplesbeforeyouputtheminafreezertopreventfromdegradingsamplesfromrepeatedfreeze-thawcycles. Q.Mysampleformedpreciptationafteraddingaceticacid.HowcanIpreventthis? A.Youcanstillmeasurereadingswithoutaddingaceticacid. Q.Theprotocolrecommendsmeasuringtheabsorbanceat490nm.Is485nmcloseenough? A.Itisokaslongasyousetuptheabsorbancebetween470and490nm. Q.WillL-LactateassaymeasureD-Lactate? A.No,itwont. Q.CanIuseatransparentwall96-wellplatewithLactateKit? A.Yes,youcan. Q.CanEthanolinterferewiththeenzyme? A.Theenzymecantolerateupto20%ofethanolwithoutaffectingit’senzymaticactivitywithL-Lactate. SamVandeVelde,MeghanF.Hogan,andMarcMontminy.:mTORlinksincretinsignalingtoHIFinductioninpancreaticbetacells.PNAS,Oct2011;108:16876-16882. JunMifune,KatrinGrage,andBerndH.A.Rehm.:ProductionofFunctionalizedBiopolyesterGranulesbyRecombinantLactococcuslactis.Appl.Envir.Microbiol.,Jul2009;75:4668-4675. DragutinJ.Savic,WilliamM.McShan,andWilliamM.McShan.:Long-termsurvivalofStreptococcuspyogenesinrichmediaispH-dependent.Microbiology,Jun2012;158:1428-1436 KristinA.Anderson,FuminLin,ThomasJ.Ribar,RobertD.Stevens,MichaelJ.Muehlbauer,ChristopherB.Newgard,andAnthonyR.Means.:DeletionofCaMKK2fromtheLiverLowersBloodGlucoseandImprovesWhole-BodyGlucoseToleranceintheMouse.Mol.Endocrinol.,Feb2012;26:281-291. JaninaP.Lewis,DivyaIyer,andCeciliaAnaya-Bergman.:AdaptationofPorphyromonasgingivalistomicroaerophilicconditionsinvolvesincreasedconsumptionofformateandreducedutilizationoflactate.Microbiology,Nov2009;155:3758-3774. AmparoWolf,SameerAgnihotri,JohannMicallef,JoydeepMukherjee,NesrinSabha,RobCairns,CynthiaHawkins,andAbhijitGuha.:Hexokinase2isakeymediatorofaerobicglycolysisandpromotestumorgrowthinhumanglioblastomamultiforme.J.Exp.Med.,Feb2011;208:313-326. InnaSerganova,AsifRizwan,XiaohuiNi,SunithaB.Thakur,JelenaVider,JamesRussell,RonaldBlasberg,andJasonA.Koutcher.:MetabolicImaging:ALinkbetweenLactateDehydrogenaseA,Lactate,andTumorPhenotype.Clin.CancerRes.,Oct2011;17:6250-6261. ChangluLiu,JiejunWu,JessicaZhu,ChesterKuei,JingxueYu,JonathanShelton,StevenW.Sutton,XiaorongLi,SuJinYun,TaranehMirzadegan,CurtMazur,FredrikKamme,andTimothyW.Lovenberg.:METABOLISM,REGULATION,ANDSIGNALING:LactateInhibitsLipolysisinFatCellsthroughActivationofanOrphanG-protein-coupledReceptor,GPR81.J.Biol.Chem.,Jan2009;284:2811-2822. MarinaOstroukhova,NicholasGoplen,MdZunayetKarim,LidiaMichalec,LeiGuo,QiaolingLiang,andRafeulAlam.:Theroleoflow-levellactateproductioninairwayinflammationinasthma.AmJPhysiolLungCellMolPhysiol,Feb2012;302:L300-L307. Zhuang,Yongxian,DanielKChan,andWKeithMiskimins.“PreventingFeedbackActivationofGlycolyticATPProductionEnhancesMetforminCytotoxicityinBreastCancerCellsWhenOxidativePhosphorylationIsInhibited.”Cancer&Metabolism2.Suppl1(2014):P89.PMC.Web.6Oct.2015. Lezi,E.etal.“EffectofExerciseonMouseLiverandBrainBioenergeticInfrastructures.”Experimentalphysiology98.1(2013):207–219.PMC.Web.6Oct.2015. YongxianZhuang,DanielK.Chan,AllisonB.Haugrud,W.KeithMiskimins.MechanismsbyWhichLowGlucoseEnhancestheCytotoxicityofMetformintoCancerCellsBothInVitroandInVivo.September25,2014DOI:10.1371/journal.pone.0108444 BorgesFT1,MeloSA,ÖzdemirBC,KatoN,RevueltaI,MillerCA,GattoneVH2nd,LeBleuVS,KalluriR.TGF-β1-containingexosomesfrominjuredepithelialcellsactivatefibroblaststoinitiatetissueregenerativeresponsesandfibrosis.JAmSocNephrol.2013Feb;24(3):385-92.doi:10.1681/ASN.2012101031.Epub2012Dec28. Everest Biotech自2000年以来，我们一直是抗肽和抗原亲和纯化山羊多克隆抗体的专家。我们的抗体具有100％的满意保证-它们可在您的实验室或您的退款中发挥作用。 LifeSpan BioSciences，Inc.收购了Everest Biotech。 在此处查找使用Everest抗体的最新出版物盖玻片 用于悬浮细胞免疫荧光 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
