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Everest Biotech/Glucose Assay Kit I (5ml)-100 Assays w/Reader Plate(1x)/120003100P/(5ml) - 100 assays w/Reader Plate (1x188bio精品生物—专注于实验室精品爆款的电商平台 - 蚂蚁淘旗下精选188款生物医学科研用品
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Everest Biotech/Glucose Assay Kit I (5ml)-100 Assays w/Reader Plate(1x)/120003100P/(5ml) - 100 assays w/Reader Plate (1x


SampleTyple

Plasma,Serum,Cellculturesupernatant,otherbodyfluid

Contents



Method

Colorimetricmethodat490nm

StandardCurve

Sensitivity


8µM-400µM

ReactionTime

15minutes

Applications

ForBIOLOGicalresearch:
Glucosemeasurementinbiologicalsamples
Fordrug/pharmresearch:
DruginfluenceonGlucosemetabolism

Storage

-80°C

Notes

Materialsneededbutnotsupplied

Aplatereadercapableofmeasuringabsorbanceat490nm

AdjustablePipettesandarepeatpipettor

Distilledwater(milliQorHPLC-grade)

0.5MAceticAcid

Clearflatbottom96-wellplatesifnotincludedinthekitpurchased

ShippingIcepacks

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
Serum,Plasma,otherbodyfluid,orcellculturesupernatantcanbemeasureddirectlybyaseriesofdilutionsofthesample(1/2;1/4;1/8;.…..)toensurethereADIngsarewithinthestandardcurve.YoursamplescanbedilutedwithdH2O.

SolidSamples
Solidsamples,suchastissues,canbefirsthomogenizedandextractedwithethanol(80%)withatissues/Ethanolratioof1:8(1hrat4°C)followedbycentrifugationat10,000xg.Theclearsupernatantsthencanbemeasureddirectlybyaseriesofdilutionsofthesample(1/2;1/4;1/8;.…..)toensurethereadingsarewithinthestandardcurve.YoursamplescanbedilutedwithdH2O.

AddingSamples
Add50μlofsamplestoeachwell.
Werecommendthatsamplesbeassayedinduplicate.

Note:
Ifpreparedsamplesarenotassayedthesameday,storethesamplesat-80?C.Thesampleswillbestableforonemonthwhilestoredat-80?C.Forfrozensamples,dilutionsofsamplesmustbedonerightbeforeassaying.

2.StandardCurvePrepation

WerecommendthatGlucoseStandardsbeassayedinduplicate.Astandardcurvehastoberunineachassay.
Add50µl,40µl,30µl,20µl,10µl,5µl,1µl,and0µlofdilutedGlucoseStandardtoeachwell.Thenadjustvolumeto50μl/wellwithdH2O.

3.Performtheassay
a.Add50µlofGlucoseAssaySolutiontoeachwellcontainingtheGlucoseStandardsandtestsamples.

b.Incubatefor15minsat37°Cincubator.Note:PleasedonotuseCO2incubator.

c.Stopthereactionbyadding50μlof0.5Maceticacidperwellfollowedbybriefgentleagitation.Note:Eliminateanyairbubblespresentinthewellsusinganeedlepriortomeasurement.

d.Measuretheabsorbanceat490nmusingamicroplatereader.

4.Calculation

a.AveragetheOD490nmvaluesofreplicatewellsofeachGlucosestandard,testsamples,andblank.Inordertogetthecorrectedabsorbance,subtracttheaverageOD490nmvalueoftheblankfromtheaveragedOD490nmvaluesfromallstandardsandsamples.

b.MakeastandardcurvebyplottingOD490nmvaluesfromeachglucosestandardsasafunctionofGlucoseconcentration.Thiscanbedonewithexcelspreadsheet.CalculatetheconcentrationofGlucoseinsamplesusingtheequationobtainedfromthelinearregressionofthestandardcurve.

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

4.FIRSTAIDMEASURES

4.1Descriptionoffirstaidmeasures

Eyecontact

Checkforandremovecontactlensesandflushwithcopiousamountsofwater;assureadequateflushingbyseparatingtheeyelidswithfingers;callaphysician

SkinContact

Flushwithcopiousamountsofwater;removecontaminatedclothingandshoes;callaphysician

Inhalation

Removetofreshair.Ifnotbreathinggiveartificialrespiration

Ingestion

Ifswallowed,nevergiveanythingbymouthtoanunconsciousperson.Rinsemouthwithwater.

4.2Mostimportantsymptomsandeffects,bothacuteanddelayed

Noinformationavailable

4.3Indicationofanyimmediatemedicalattentionandspecialtreatmentneeded.

Noinformationavailable

5.FIREFIGHTINGMEASURES

5.1Extinguishingmedia

Suitableextinguishingmedia

Waterspray,alcohol-resistantfoam,drychemicalorcarbondioxide

5.2Specialhazardsarisingfromthesubstanceormixture

Noinformationavailable

5.3Adviceforfirefighters

Wearself-containedbreathingapparatusandprotectiveclothingtopreventcontactwithskinandeyes.

6.ACCIDENTALRELEASEMEASURES

6.1Personalprecautions,protectiveequipmentandemergencyprocedures

Usefullpersonalprotectiveequipment.Evacuatepersonneltosafeareas.Avoidbreathingvapors,mistorgas.

6.2Environmentalprecautions

Preventfurtherleakageorspillage.Preventproductfromenteringdrain.

6.3Methodsandmaterialforcontainmentandcleaningup

Keepinsuitable,closedcontainersfordisposal

7.HANDLINGANDSTORAGE

7.1Precautionsforsafehandling

Avoidskin/eyecontact.Useprotectiveequipmentasneeded.Washcontaminatedclothingbeforereuse.

7.2Conditionsforsafestorage,includinganyincompatibilities

Keepcontainertightlyclosed.Storeat-80?C.

7.3Specificenduse(s)

Nodataavailable.

8.EXPOSURECONTROLS/PERSONALINFORMATION

8.1ControlParameters

Thisproductcontainsnohazardousmaterialswithoccupationalexposurelimits.

8.2Exposurecontrols

Engineeringcontrols

Provideshowerandeyewashstation

Personalprotectiveequipment

GlucoseAssaySolution

Components

CAS#

Potassiumphosphatemonobasic

7778-77-0

Potassiumphosphatedibasic

7758-11-4

GlucoseOxidase

9001-37-0

GlucoseAssayStandard

Components

CAS#

SodiumD-Glucose

50-99-7

Potassiumphosphatemonobasic

7778-77-0

Potassiumphosphatedibasic

7758-11-4

Water

7732-18-5









EyeProtection

Wearsafetygoggles

SkinProtection

Wearprotectiveclothing

HandProtection

Wearprotectivegloves

Respiratoryprotection

WearNIOSH/MSHAapprovedrespirators

9.PHYSICALANDCHEMICALPROPERTIES

9.1Informationonbasicphysicalandchemicalproperties

Appearance

Clear,colorlessliquid

pH

7.0

Meltingpoint

Nodataavailable

Freezingpoint

Nodataavailable

Boilingpoint

Nodataavailable

Density

Nodataavailable

Watersolubility

Soluble

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
Non-Hazardousfortransport:thissubstanceisconsideredtobenon-hazardousfortransport
IATA:

Propershippingname:none
Non-Hazardousfortransport:thissubstanceisconsideredtobenon-hazardousfortransport

ADR/RID

Propershippingname:none
Non-Hazardousfortransport:thissubstanceisconsideredtobenon-hazardousfortransport

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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