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Everest Biotech/Galactose Assay Kit 100 assays/1200091002/100 assays

SampleTyple

Plasma,Serum,Cellculturesupernatant,otherbodyfluid

Components

Method

Colorimetricmethodat570nm

StandardCurve

Sensitivity

20µM-1000µM

ReactionTime

30minutes

Applications

ForBIOLOGicalresearch:
Galactosemeasurementinbiologicalsamples
Fordrug/pharmresearch:
DruginfluenceonGalactosemetabolism

Storage

-80°C

Notes

Materialneededbutnotsupplied

Aplatereadercapableofmeasuringabsorbanceat570nm

AdjustablePipettesandarepeatpipettor

Distilledwater(milliQorHPLC-grade)

ClearFlatbottom96-wellplatesifnotincludedinthekitpurchased

ShippingIcepacks

ReagentPreparation

Note:Allreagentsarefrozen.Werecommendyouspinsmallvialsbeforeopening.


1.GalactoseAssayBuffer(10x)

Note:Pleasedon’tdodilutionintheprovidedGalactoseAssayBufferbottleitself.


Thebottlecontains10mlof10xGalactoseAssayBuffer.Theassaybuffermustbeequilibratedtoroomtemperature.Pleasemakeappropriateamountof1xGalctoseAssayBufferfortheassay,forexample,bydiluting1mlof10xGalactoseAssayBufferwith9mlofdistilledwater.Prepareadditionaldilutedassaybufferifneeded.Storeat-80?C.

2.GalactoseStandard(10x)

Note:Pleasedon’tdodilutionintheprovidedGalactoseStandardvialitself.


Thevialcontains100µlof10mMGalactoseStandard.Thestandardmustbeequilibratedtoroomtemperaturebeforeuse.Dilute100µlof10mMGalactoseStandardwith900µlofdilutedGalactoseAssayBuffertopreparea1mMGalactoseStandard.1mlofdilutedGalctoseStandardisenoughformaking3standardcurvesifassayedinduplicate.Storeat-80?C.

3.AssayProbe

Thevialcontains45μlofAssayProbe.Theassayprobemustbeequilibratedtoroomtemperaturebeforeuse.Itissufficientfor100wells.Protectfromlight.Storeat-80?C.

4.GalactoseEnzymeMix

Thevialcontains35μlofGalctoseEnzymeMix.Theenzymemixmustbethawedonicebeforeuse.Itissufficientfor100wells.Storeat-80?C.

Protocol

1.SamplePreparation

Serum,plasma,urine,otherbodyfluidscanbemeasureddirectlybyaseriesofdilutionsofthesample(1/2;1/4;1/8;…)toensurethereADIngsarewithinthestandardcurverange.YoursamplescanbedilutedwithdH2OordilutedGalactoseAssayBuffer.

Add50μlofsamplestoeachwell.
Werecommendthatsamplesbeassayedinduplicate.

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

2.StandardCurvePrepation

Add50μl,40μl,30μl,20μl,10μl,5μl,1μl,and0μlofdilutedGalactoseStandardtoeachwell,thenadjustvolumeto50μl/wellwithdilutedGalactoseAssayBuffer.
WerecommendthatGalactoseStandardsbeassayedinduplicate.Astandardcurvehastoberunineachassay.

3.ReactionSolutionPreparation
50μlReactionMixisrequiredforeachwelland1mlofReactionSolutionispreparedwiththefollowings:
986µlGalactoseAssayBuffer(1x)

6µlGalactoseAssayEnzymeMix
8µlAssayProbe


1mlofReactionSolutionisenoughfor~20wells.

PrepareappropriateamountofReactionSolutionforthenumberofassaystobeperformed.

Note:PleasedonotkeepanyleftoverReactionSolutionasithastobemadefreshrightbeforeyourunassays.IfoldReactionSolutionisused,assayswillnotwork.

4.Performtheassay

a.Mixthereactionsolutionwellandadd50μlofthereactionsolutiontoeachwellcontainingtheGalactosestandardsandtestsamples.

b.Incubatethereactionfor30minutesina37?Cincubator.

c.Measureabsorbanceat570nminamicroplatereader.

5.Calculation

a.AveragetheOD570nmvaluesofreplicatewellsofeachGalactoseStandard,testsamples,andblank.Then,correctbackgroundbysubtractingthevaluederivedfromtheblankfromallsamplereadings.

b.MakeastandardcurvebyplottingOD570nmvaluesfromeachGalactoseStandardsasafunctionofGalactoseconcentration.Thiscanbedonewithexcelspreadsheet.

c.CalculatethevalueofGalactoseinsamplesusingtheequationobtainedfromthelinearregressionofthestandardcurve.

Galactose(μM)=[(Correctedabsorbance)-(y-intercept)]/Slope

MaterialSafetyDataSheet

Date:Oct26,2015

1.PRODUCTANDCOMPANYIDENTIFICATION

1.1Productidentifier

Productname:GalactoseAssayKit

CatalogNo:1200091002,120009100P

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

GalactoseAssayBuffer

GalactoseAssayEnzymeMix

Components

CAS#

Components

CAS#

PotassiumPhosphateMonobasic

7778-77-0

PotassiumPhosphateMonobasic

7778-77-0

PotassiumPhosphateDibasic

7758-11-4

PotassiumPhosphateDibasic

7758-11-4

Water

7732-18-5

GalactoseOxidase

9028-79-9

GalactoseStandard

AssayProbe

Components

CAS#

Components

CAS#

SodiumD-Galactose

59-23-4

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

GalactoseAssayBuffer

Appearance

Liquid,colorless

pH

7.0

Meltingpoint

Nodataavailable

Freezingpoint

Nodataavailable

Boilingpoint

Nodataavailable

Density

Nodataavailable

Watersolubility

Soluble

GalactoseStandard

Appearance

Liquid,colorless

pH

7.0

Meltingpoint

Nodataavailable

Freezingpoint

Nodataavailable

Boilingpoint

Nodataavailable

Density

Nodataavailable

Watersolubility

Soluble

GalactoseAssayEnzymeMix

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.


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