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主营:研究并生产荧光和发光探针,信号转导研究的试剂
℡ 4000-520-616
℡ 4000-520-616
AAT Bioquest/Amplite™ Colorimetric Ammonia Quantitation Kit *Blue Color*/10059/200 Tests
产品编号:10059
市  场 价:¥56560.00
场      地:美国(厂家直采)
联系QQ:1570468124
电话号码:4000-520-616
邮      箱: info@ebiomall.com
美  元  价:$2828.00
品      牌: AAT Bioquest
公      司:AAT Bioquest
公司分类:
AAT Bioquest/Amplite™ Colorimetric Ammonia Quantitation Kit *Blue Color*/10059/200 Tests
商品介绍
Overview
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Ex/Em(nm)650/None
MWN/A
CAS#N/A
SolventN/A
StorageF/D/L
CategorySmallMoleculeDetection
DiagnosticMolecules
RelatedCellMetabolism
BiochemicalAssays
Ammoniaisanimportantsourceofnitrogenforlivingsystems.Itissynthesizedthroughaminoacidmetabolismandistoxicwhenpresentathighconcentrations.Itisproducedinliverandconvertedtoureathroughtheureacycle.Elevatedlevelsofammoniaintheblood(hyperammonemia)havebeenfoundinliverdysfunction(cirrhosis),whilehypoammonemiahasbeenassociatedwithdefectsintheureacycleenzymes(e.g.ornithinetranscarbamylase).Thedeterminationofammoniaisveryusefultestinclinicallaboratorytomonitorhealthstatus.OurAmplite™ColorimetricAmmoniaAssayKitprovidesasimpleandsensitivecolorimetricmethodforthequantitationofammoniaconcentrationinfoodsandBIOLOGicalsamplessuchasserum,plasmaandurine,etc.Theassayisbasedonanenzyme-coupledreactionofammoniaintheassaybuffer,andfinallyproducesabluecoloredproduct.Theintensityofcolorproducedisproportionaltotheconcentrationofammoniainthesample,whichcanbemeasuredcolorimetricallyat660-670nm.ThisAmplite™ColorimetricAmmoniaAssayKitprovidesasimpleassaytodetectammonia.Theassaycanbeperformedinaconvenient96-wellor384-wellmicrotiter-plateformatandeasilyadaptedtoautomationwithoutaseparationstep.

Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.

1.      PrepareserialdilutionsofAmmoniumChloride(0to1mM)solutions:

1.1   Add1μLof1.0MAmmoniumChlorideStandard(ComponentC)to999µLDPBStogenerate1.0mMstandard

ammoniumchloridesolution.

1.2   Take300μLof1.0mMstandardtoperform1:3serialdilutionstoget300,100,30,10,3,1,and0µMstandard 

       ammoniumchloridesolutions.

1.3   AddAmmoniumChlorideStandardsandammoniacontainingtestsamplesintoa96-wellclearbottommicroplateasdescribedinTables1and2.

Table1.Layoutofstandardsandtestsamplesinaclearbottom96-wellmicroplate:

BL

BL

TS

TS

….

….

 

 

 

 

 

 

AS1

AS1

….

….

….

….

 

 

 

 

 

 

AS2

AS2

 

 

 

 

 

 

 

 

 

 

AS3

AS3

 

 

 

 

 

 

 

 

 

 

AS4

AS4

 

 

 

 

 

 

 

 

 

 

AS5

AS5

 

 

 

 

 

 

 

 

 

 

AS6

AS6

 

 

 

 

 

 

 

 

 

 

AS7

AS7

 

 

 

 

 

 

 

 

 

 

Note:AS=AmmoniumChlorideStandards,BL=BlankControl,TS=TestSamples.

 

Table2.Reagentcompositionforeachwell:

AmmoniumChlorideStandard

BlankControl

TestSample

Serialdilutions*:50μL

DPBS:50μL

50μL

*Note:Addtheseriallydilutedammoniumchloridestandardsfrom1to1000µMintowellsfromAS1toAS7induplicate.

2.      RunAmmoniumChlorideAssay:

2.1   Add50μLofAssayBufferI(ComponentA)toeachwelloftheAmmoniumChlorideStandard,blankcontrol,andtestsamples(seeStep1.3)sothatthetotalassayvolumeis100µL/well.

Note:Fora384-wellplate,add25μLsample,25μLofAssayBufferIperwell.

2.2   Incubatethereactionfor5minutesatroomtemperature.

2.3   Add50μLofAssayBufferII(ComponentB)toeachwellsothatthetotalassayvolumeis150µL/well.

Note:Fora384-wellplate,add25μLAssayBufferII(ComponentB)toeachwell.

2.4   Incubateatroomtemperaturefor30-60minutes,andmonitortheabsorbanceincreaseat660-670nmusinganabsorbancemicroplatereader.

Note:ThecolorturnstoyellowafterAssayBufferII(ComponentB)isadded,andthewellswithAmmoniumChlorideStandardorsampleswillshowbluishgreencolorafterincubation.Theintensityofthecolorwillreachthemaximumin30-60minutes.

References&Citations
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1.   LohTP,TanKM,ChewS,ChanDS.(2013)Highpleuralammonianegativelyinterfereswiththemeasurementofadenosinedeaminaseactivity.BMJCaseRep,2013.

2.   BakLK,WaagepetersenHS,SorensenM,OttP,VilstrupH,KeidingS,SchousboeA.(2013)Roleofbranchedchainaminoacidsincerebralammoniahomeostasisrelatedtohepaticencephalopathy.MetabBrainDis.

3.   TranahTH,VijayGK,RyanJM,ShawcrossDL.(2013)Systemicinflammationandammoniainhepaticencephalopathy.MetabBrainDis,28,1.

4.   HenryRP,LucuC,OnkenH,WeihrauchD.(2012)Multiplefunctionsofthecrustaceangill:osmotic/ionicregulation,acid-basebalance,ammoniaexcretion,andbioaccumulationoftoxicmetals.FrontPhysiol,3,431.

5.   RothmanDL,DeFeyterHM,MaciejewskiPK,BeharKL.(2012)Isthereinvivoevidenceforaminoacidshuttlescarryingammoniafromneuronstoastrocytes?NeuRochemRes,37,2597.

6.   StahlDA,delaTorreJR.(2012)Physiologyanddiversityofammonia-oxidizingarchaea.AnnuRevMicrobiol,66,83.

7.   ShenJP,ZhangLM,DiHJ,HeJZ.(2012)Areviewofammonia-oxidizingbacteriaandarchaeainChinesesoils.FrontMicrobiol,3,296.

8.   WrightPA,WoodCM.(2012)Seventhingsfishknowaboutammoniaandwedon"t.RespirPhysiolNeurobiol,184,231.

9.   RoseCF.(2012)Ammonia-loweringstrategiesforthetreatmentofhepaticencephalopathy.ClinPharmacolTher,92,321.

10.   deVilliersM,PuthanVeetilV,RajH,deVilliersJ,PoelarendsGJ.(2012)Catalyticmechanismsandbiocatalyticapplicationsofaspartateandmethylaspartateammonialyases.ACSChemBiol,7,1618.


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