1.D-LactateStandards

Thevialcontains1mlof3mMD-LactateStandard.Thestandardmustbeequilibratedtoroomtemperaturebeforeuse.1mlofthestandardisenoughformaking3standardcurvesifassayedinduplicate.Storeat-80˚C.

2.D-LactateAssaySolution

Thesolutioncontainsenzymesthatarelightsensitive.Thesolutionmustbethawedonicebeforeuse.Besttoaliquottheamountneededanduseitalltopreventthawing/freezingcycles.Freezeandstorealiquotsat-80oC

1.SamplePreparation

Serum/Plasma/otherbodyfluid/cellculturesupernatant

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

YoursamplescanbedilutedwithdH2O.

Note:ifsamples(suchashemolyzedserum/plasmaorcellculturemediumwhichcontainFBS)containhighleveloflactatedehydrogenasecapableofconvertinglactatetopyruvate,itisimportantforthesamplestobedeproteinated.

SolidSamples

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

AddingSamples

Add50μlofsamplestoeachwell.

Werecommendthatsamplesbeassayedinduplicate.

Ifpreparedsamplesarenotassayedthesameday,storethesamplesat-80˚C.Ifsamplesneedtobedeproteinated,makesuretodeproteinizethesamplespriortostoringinthefreezer.Thedeproteinatedsampleswillbestableforonemonthwhilestoredat-80˚C.Forfrozensamples,dilutionsofsamplesmustbedonerightbeforeassaying.

2.StandardCurvePreparation

WerecommendthatD-LactateStandardsbeassayedinduplicate.Astandardcurvehastoberunineachassay.
Add50µl,40µl,30µl,20µl,10µl,5µl,1µl,and0µlofD-LactateStandardtoeachwell.Thenadjustvolumeto50μl/wellwithdH2O.

3.PerformtheAssay

a.Add50µlofD-LactateAssaySolutiontoeachwellcontainingtheD-Lactatestandardsandtestsamples.

b.Incubatefor30minsat37°Cincubator.Note:PleasedonotuseCO2incubator.

c.Stopthereactionbyadding50μlof0.5Maceticacidperwellfollowedbybriefgentleagitation.Note:Eliminateanyairbubblespresentinthewellsusinganeedlepriortomeasurement.

d.Measuretheabsorbanceat490nmusingamicroplatereader.

4.Calculation

a.AveragetheOD490nmvaluesofreplicatewellsofeachD-Lactatestandard,testsamples,andblank.Inordertogetthecorrectedabsorbance,subtracttheaverageOD490nmvalueoftheblankfromtheaverageOD490nmvaluesfromallstandardsandsamples.

b.MakeastandardcurvebyplottingOD490nmvaluesfromeachD-LactatestandardsasafunctionofD-Lactateconcentration.Thiscanbedonewithexcelspreadsheet.CalculatethevalueofD-Lactateinsamplesusingtheequationobtainedfromthelinearregressionofthestandardcurve.

D-Lactate(µM)=[(Correctedabsorbance)-(y-intercept)]/slope

MaterialSafetyDataSheet

Date:Oct26,2015

1.PRODUCTANDCOMPANYIDENTIFICATION

1.1Productidentifier

Productname:D-LactateAssayKit

CatalogNo:1200021002,120002100A,120002100P,1200022002,120002200A,12002200P,1200024002,120002400A,120002400P

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;callaphysician

Ingestion

Ifswallowed,nevergiveanythingbymouthtoanunconsciousperson.Rinsemouthwithwater;callaphysician

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

	D-lactateAssaySolution
	Components		CAS#
	DMSO		67-68-5
	D-LDH		9028-36-8
	Tris-HCl,pH7.5		N/A
	Water		7732-18-5
	D-LactateAssayStandard
	Components		CAS#
	SodiumD-Lactate		920-49-0
	Tris-HCl,pH7.5		N/A
	Water		7732-18-5

EyeProtection	Wearsafetygoggles
SkinProtection	Wearprotectiveclothing
HandProtection	Wearprotectivegloves
Respiratoryprotection	WearNIOSH/MSHAapprovedrespirators

9.PHYSICALANDCHEMICALPROPERTIES

9.1Informationonbasicphysicalandchemicalproperties

L-LactateAssaySolution

Appearance	Liquid,yellowish
pH	7.5
Meltingpoint	Nodataavailable
Freezingpoint	Nodataavailable
Boilingpoint	Nodataavailable
Density	Nodataavailable
Watersolubility	Soluble

L-LactateStandard

Appearance	Liquid,colorless
pH	7.5
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.