产品资料

Everest Biotech/D-Lactate Assay Kit (10 ml)-200 Assays- Assay Solution ONLY/120002200A/(10 ml)-200 Assays- Assay Soluti

SampleTyple	Plasma,Serum,Cellculturesupernatant,otherbodyfluid
Contents
Method	Colorimetricmethodat490nm
StandardCurve
Sensitivity	60µM-3000µM
ReactionTime	30minutes
Applications	ForBIOLOGicalresearch: D-Lactatemeasurementinbiologicalsamples Fordrug/pharmresearch: DruginfluenceonD-Lactatemetabolism
Storage	-80°C
Notes	Materialsneededbutnotsupplied Aplatereadercapableofmeasuringabsorbancebetween470-490nm AdjustablePipettesandarepeatpipettor Asourceofpurewater;glassdistilledwaterofHPLC-gradewaterissufficient 0.5MAceticAcid Clearflatbottom96-wellplatesifnotincludedinthekitpurchased
Shipping	Icepacks

ReagentPreparation

1.D-LactateStandards

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

2.D-LactateAssaySolution

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

AssayProtocol

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.

Note:

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.

Q.whattypeofmediumshouldIuseformakingculturedcellsforLactateassay?

A.Pleaseusephenolredfreemedium.Pleasedonotusephenolredmediumsincephenolredwouldaffectabsorbancereadings.

Q.WhatenzymesisusedinLactateAssay?

A.ItisLDH.

Q.CanyoutellmethecompositionsofyourLactateAssay?

A.Unfortunatelyitisproprietary.

Q.WhattypeofanticoagulantshouldIuseformysamples?

A.Ifyouarecollectingbloodformeasuringlactatefromserum,youdonothavetocollectbloodusingananticoagulant.Ifyouarecollectingbloodformeasuringlactatefromblood,youcanuseheparinorEDTA.

Q.HowdoIknowwhetherIneedtouseL-LactateorD-Lactateassaykit?

A.L-Lactateassaykitsarecommonlyusedformeasuringlactatelevelinanimalcells.

D-lactateassaykitsarecommonlyusedformeasuringlactatelevelinbacteriacells.

Q.DoIneeedtomakeastandardcurveeverytime?

A.Yesitisnecessarysinceyoucalculateyoursampleconcentrationbasedonthestandardcurve.PleasedonotusetherepresentativeLactatestandardcurveintheprotocoltocalculateconcentrationsofyoursamples.

Q.CanIusefluorescencespectroscopytomeasurereadingsforLactateassaykits?

No,thekitdoesnotworkwithafluorescencereadersincethekitemployscolorimetricassays.

Q.HowDoImeasureLactatefromculturedcells?

A.Youcangrowcellsinphenolred-freemedium,thenyoucanextractcellswith80%ethanol.

Q.CanIusefrozensamples?

Althoughitisbettertousefreshsamplesforassays,youcanusefrozensamples.Ifyouarenotusingfrozensamplesinonce,pleasemakealiquotsofyoursamplesbeforeyouputtheminafreezertopreventfromdegradingsamplesfromrepeatedfreeze-thawcycles.

Q.Mysampleformedprecipitationafteraddingaceticacid.HowcanIpreventthis?

A.Youcanstillmeasurereadingswithoutaddingaceticacid.

Q.Theprotocolrecommendsmeasuringtheabsorbanceat490nm.Is485nmcloseenough?

A.Itisokaslongasyousetuptheabsorbancebetween470and490nm.

Q.WillD-LactateassaykitmeasureL-Lactate?

A.Noitwont

Neal-McKinneyJM,LuX,DuongT,LarsonCL,CallDR,etal.(2012)ProductionofOrganicAcidsbyProbioticLactobacilliCanBeUsedtoReducePathogenLoadinPoultry.PLoSONE7(9):e43928.doi:10.1371/journal.pone.0043928

IwataniS,NagashimaH,ReddyR,ShiotaS,GrahamDY,YamaokaY.IdentificationofthegenesthatcontributetolactateutilizationinHelicobacterpylori.PLosOne.2014Jul31;9(7):e103506.doi:10.1371/journal.pone.0103506.eCollection2014.

ApontesP,LiuZ,SuK,BenardO,YounDY,LiX,LiQ,MirzaRH,BastieCC,JelicksLA,PessinJE,MuzumdarRH,SauveAA,ChiY.Mangiferinstimulatescarbohydrateoxidationandprotectsagainstmetabolicdisordersinducedbyhigh-fatdiets.Diabetes.2014Nov;63(11):3626-36.doi:10.2337/db14-0006.Epub2014May21.

MeyerRE,WhitleyJT,MorrowWEM,etal.Effectofphysicalandinhaledeuthanasiamethodsonhormonalmeasuresofstressinpigs.JSwineHealthProd.2013;21(5):261–269.

BurkhartRA1,PinedaDM,ChandSN,RomeoC,LondinER,KarolyED,CozzitortoJA,RigoutsosI,YeoCJ,BrodyJR,WinterJM.HuRisapost-transcriptionalregulatorofcoremetabolicenzymesinpancreaticcancer.RNABiol.2013Aug;10(8):1312-23.doi:10.4161/rna.25274.Epub2013Jun13.

KarthiyainiDamodharan,SasikumarArunachalamPalaniyandi,SeungHwanYang,Joo-WonSuh.InvitroprobioticcharacterizationofLactobacillusstrainsfromfermentedradishandtheiranti-adherenceactivityagainstentericpathogens.CanadianJournalofMicrobiology,Publishedontheweb10August2015,10.1139/cjm-2015-0311

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Everest Biotech/D-Lactate Assay Kit (10 ml)-200 Assays- Assay Solution ONLY/120002200A/(10 ml)-200 Assays- Assay Soluti