SampleTyple | Plasma,Serum,Cellculturesupernatant,otherbodyfluid | |||||||
Components | ||||||||
Method | Colorimetricmethodat570nm | |||||||
StandardCurve | ||||||||
Sensitivity | 4µM-200µM | |||||||
ReactionTime | 30minutes | |||||||
Applications | ForBIOLOGicalresearch: | |||||||
Storage | -80°C | |||||||
| Notes | Materialneededbutnotsupplied Aplatereadercapableofmeasuringabsorbanceat570nm AdjustablePipettesandarepeatpipettor Distilledwater(milliQorHPLC-grade) ClearFlatbottom96-wellplatesifnotincludedinthekitpurchased | |||||||
| Shipping | Icepacks | |||||||
ReagentPreparation Note:Allreagentsarefrozen.Werecommendyouspinsmallvialsbeforeopening. 1.GlucoseAssayBuffer(10x) Note:Pleasedon’tdodilutionintheprovidedGlucoseAssayBufferbottleitself. Thebottlecontains10mlof10xGlucoseAssayBuffer.Theassaybuffermustbeequilibratedtoroomtemperature.Pleasemakeappropriateamountof1xGlucoseAssayBufferfortheassay,forexample,bydiluting1mlof10xGlucoseAssayBufferwith9mlofdistilledwater.Prepareadditionaldilutedassaybufferifneeded.Storeat-80?C. 2.GlucoseStandard(10x) Note:Pleasedon’tdodilutionintheprovidedGlucoseStandardvialitself. Thevialcontains100µlof2mMGlucoseStandard.Thestandardmustbeequilibratedtoroomtemperaturebeforeuse.Dilute100µlof2mMGlucoseStandardwith900µlofdilutedGlucoseAssayBuffertopreparea200μMGlucoseStandard.1mlofdilutedGlucoseStandardisenoughformaking3standardcurvesifassayedinduplicate.Storeat-80?C. 3.AssayProbe Thevialcontains45μlofAssayProbe.Theassayprobemustbeequilibratedtoroomtemperaturebeforeuse.Itissufficientfor100wells.Protectfromlight.Storeat-80?C. 4.GlucoseEnzymeMix Thevialcontains70μlofGlucoseEnzymeMix.Theenzymemixmustbethawedonicebeforeuse.Itissufficientfor100wells.Storeat-80?C. | ||||||||
Protocol 1.SamplePreparation Blood(serum/plasma),urine,otherbodyfluidscanbemeasureddirectlybyaseriesofdilutionsofthesample(1/2;1/4;1/8;…)toensurethereADIngsarewithinthestandardcurverange.YoursamplescanbedilutedwithdH2OordilutedGlucoseAssayBuffer. Note: | ||||||||
2.StandardCurvePrepation Add50μl,40μl,30μl,20μl,10μl,5μl,1μl,and0μlofdilutedGlucoseStandardtoeachwell,thenadjustvolumeto50μl/wellwithdilutedGlucoseAssayBuffer. | ||||||||
3.ReactionSolutionPreparation 12µlGlucoseAssayEnzymeMix 8µlAssayProbe 1mlofReactionSolutionisenoughfor~20wells. PrepareappropriateamountofReactionSolutionforthenumberofassaystobeperformed. Note:PleasedonotkeepanyleftoverReactionSolutionasithastobemadefreshrightbeforeyourunassays.IfoldReactionSolutionisused,assayswillnotwork. | ||||||||
4.Performtheassay a.Mixthereactionsolutionwellandadd50μlofthereactionsolutiontoeachwellcontainingtheGlucosestandardsandtestsamples. | ||||||||
5.Calculation a.AveragetheOD570nmvaluesofreplicatewellsofeachGlucoseStandard,testsamples,andblank.Then,correctbackgroundbysubtractingthevaluederivedfromtheblankfromallsamplereadings. Glucose(μM)=[(Correctedabsorbance)-(y-intercept)]/Slope | ||||||||
GlucoseAssayBuffer | GlucoseAssayEnzymeMix | |||||||
Components | CAS# | Components | CAS# | |||||
PotassiumPhosphateMonobasic | 7778-77-0 | PotassiumPhosphateMonobasic | 7778-77-0 | |||||
PotassiumPhosphateDibasic | 7758-11-4 | PotassiumPhosphateDibasic | 7758-11-4 | |||||
Water | 7732-18-5 | GlucoseOxidase | 9001-37-0 | |||||
GlucoseStandard | AssayProbe | |||||||
Components | CAS# | Components | CAS# | |||||
SodiumD-Glucose | 50-99-7 | DMSO | 67-68-5 | |||||
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
GlucoseAssayBuffer
Appearance | Liquid,colorless |
pH | 7.0 |
Meltingpoint | Nodataavailable |
Freezingpoint | Nodataavailable |
Boilingpoint | Nodataavailable |
Density | Nodataavailable |
Watersolubility | Soluble |
GlucoseStandard
Appearance | Liquid,colorless |
pH | 7.0 |
Meltingpoint | Nodataavailable |
Freezingpoint | Nodataavailable |
Boilingpoint | Nodataavailable |
Density | Nodataavailable |
Watersolubility | Soluble |
GlucoseAssayEnzymeMix
Appearance | Liquid,yellow |
pH | Nodataavailable |
Meltingpoint | Nodataavailable |
Freezingpoint | Nodataavailable |
Boilingpoint | Nodataavailable |
Density | Nodataavailable |
Watersolubility | Soluble |
AssayProbe
Appearance | Liquid,colorless |
pH | Nodataavailable |
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.
