SampleTyple | Plasma,Serum,Cellculturesupernatant,otherbodyfluid |
Components | |
Method | Colorimetricmethodat570nm |
StandardCurve | |
Sensitivity | 2µM-100µM |
ReactionTime | 30minutes |
Applications | ForBIOLOGicalresearch: |
Storage | -80°C |
| Notes | Materialneededbutnotsupplied Aplatereadercapableofmeasuringabsorbanceat570nm AdjustablePipettesandarepeatpipettor Distilledwater(milliQorHPLC-grade) ClearFlatbottom96-wellplatesifnotincludedinthekitpurchased |
| Shipping | Icepacks |
ReagentPreparation Note:Allreagentsarefrozen.Werecommendyouspinsmallvialsbeforeopening. Note:Pleasedon’tdodilutionintheprovidedL-LactateAssayBufferbottleitself. Thebottlecontains10mlof10xL-LactateAssayBuffer.Theassaybuffermustbeequilibratedtoroomtemperaturebeforeuse.Pleasemakeappropriateamountof1xL-LactateAssayBufferfortheassay,forexample,bydiluting1mlof10xL-LactateAssayBufferwith9mlofdistilledwater.Prepareadditionaldilutedassaybufferifneeded.Storeat-80?C.2.L-LactateStandard(10x) Note:Pleasedon’tdodilutionintheprovidedL-Lactatestandardvialitself. Thevialcontains100µlof1mML-LactateStandard.Thestandardmustbeequilibratedtoroomtemperaturebeforeuse.Dilute100µlof1mML-LactateStandardwith900µlofdilutedL-LactateAssayBuffertopreparea100μML-LactateStandard.1mlofdilutedstandardisenoughformaking3standardcurvesifassayedinduplicate.Storeat-80?C. 3.AssayProbe Thevialcontains45μlofAssayProbe.Itissufficientfor100wells.Theassayprobemustbeequilibratedtoroomtemperaturebeforeuse.Protectfromlight.Storeat-80?C. 4.L-LactateEnzymeMix Thevialcontains55μlofL-LactateEnzymeMix.Theenzymemixmustbethawedonicebeforeuse.Itissufficientfor100wells.Storeat-80?C. | ||||||||
Protocol 1.SamplePreparation Blood,Serum,Plasma,otherbodyfluid,orcellculturesupernatantcanbemeasureddirectlybyaseriesofdilutionsofthesample(1/2;1/4;1/8;…)toensurethereADIngsarewithinthestandardcurverange.YoursamplescanbedilutedwithdH2OordilutedL-LactateAssayBuffer.However,keepinmindifsamples(suchashemolyzedserum/plasma)containhighleveloflactatedehydrogenasecapableofconvertinglactatetopyruvate,itisimportantforthesamplestobedeproteinated. | ||||||||
2.StandardCurvePrepation Add50µl,40µl,30µl,20µl,10µl,5µl,1µl,and0µlofdilutedL-LactateStandardtoeachwell,thenadjustvolumeto50μl/wellwithdilutedL-LactateAssayBuffer. | ||||||||
3.ReactionSolutionPreparation | ||||||||
4.Performtheassay a.Mixthereactionsolutionwellandadd50μlofthereactionsolutiontoeachwellcontainingtheL-Lactatestandardsandtestsamples. | ||||||||
5.Calculation a.AveragetheOD570nmvaluesofreplicatewellsofeachL-LactateStandard,testsamples,andblank.Then,correctbackgroundbysubtractingthevaluederivedfromtheblankfromallsamplereadings. L-Lactate(μM)=[(Correctedabsorbance)-(y-intercept)]/Slope | ||||||||
L-LactateAssayBuffer | L-LactateAssayEnzymeMix | |||||||
Components | CAS# | Components | CAS# | |||||
PotassiumPhosphateMonobasic | 7778-77-0 | PotassiumPhosphateMonobasic | 7778-77-0 | |||||
PotassiumPhosphateDibasic | 7758-11-4 | PotassiumPhosphateDibasic | 7758-11-4 | |||||
Water | 7732-18-5 | LactateOxidase | 9028-72-2 | |||||
L-LactateAssayStandard | AssayProbe | |||||||
Components | CAS# | Components | CAS# | |||||
SodiumL-Lactate | 867-56-1 | 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
L-LactateAssayBuffer
Appearance | Liquid,colorless |
pH | 7.0 |
Meltingpoint | Nodataavailable |
Freezingpoint | Nodataavailable |
Boilingpoint | Nodataavailable |
Density | Nodataavailable |
Watersolubility | Soluble |
L-LactateStandard
Appearance | Liquid,colorless |
pH | 7.0 |
Meltingpoint | Nodataavailable |
Freezingpoint | Nodataavailable |
Boilingpoint | Nodataavailable |
Density | Nodataavailable |
Watersolubility | Soluble |
L-LactateAssayEnzymeMix
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.
