请使用支持JavaScript的浏览器!
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™ Fluorimetric D-Lactate Assay Kit/13810/200 Tests
产品编号:13810
市  场 价:¥85560.00
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:$4278.00
品      牌: AAT Bioquest
公      司:AAT Bioquest
公司分类:
AAT Bioquest/Amplite™ Fluorimetric D-Lactate Assay Kit/13810/200 Tests
商品介绍
Overview
PrinterFriendlyVersion

Ex/Em(nm)571/585
MWN/A
CAS#N/A
SolventN/A
StorageF/D/L
CategoryCellBIOLOGy
CellMetabolism
RelatedRedoxEnzymes
BiochemicalAssays
Lacticacidischiralandhastwoopticalisomers:L-lacticacidandD-lacticacid.Lactateisconstantlyproducedfrompyruvateviatheenzymelactatedehydrogenase(LDH)intheprocessofmetabolismandexercise.Monitoringlactatelevelsisagoodwaytoevaluatethebalancebetweentissueoxygendemandandutilizationandisusefulwhenstudyingcellularandanimalphysiology.D-lactateisnotmetabolizedbymammalsanditseliminationfromthebodydependsmainlyonrenalexcretion.D-andL-lacticacidarefoundinmanyfermentedmilkproductssuchasyoghurtandcheese,andalsoinpickledvegetables,andcuredmeatsandfish.TheD-andL-lacticacid(generatedbybacteria)isaqualityindicatoroffoods,suchasegg,milk,fruitjuiceandwine.AbnormalhighconcentrationofD-lactateinthebloodisusuallyareflectionofbacterialovergrowthinthegastrointestinaltract.AATBioquest"sAmplite™LactateAssayKits(Cat#13814and13815forL-lactateassay,andCat#13810and13811forD-lactateassay)providebothfluorescenceandabsorbance-basedmethodfordetectingeitherL-lactateorD-lactateinbiologicalsamplessuchasserum,plasma,urine,aswellasincellculturesamples.Intheenzymecoupledassay,lactateisproportionallyrelatedtoNADH,whichisspecificallymonitoredbyafluorogenicNADHsensor.ThesignalcanbereadbyafluorescencemicroplatereaderatEx/Em=540nm/590nm.WiththisFluorimetricAmplite™D-LactateAssayKit,wewereabletodetectaslittleas1.4µMD-lactateina100µLreactionvolume.

Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.

1.PrepareNADstocksolution(100X):

Add100µLofH2OintothevialofNAD(ComponentC)tomake100XNADstocksolution.

 

2.PrepareD-lactatestocksolution:

Add200µLofH2Oor1xPBSbufferintothevialofD-lactateStandard(ComponentD)tomake100mMD-lacatestandardsolution.

Note:TheunusedD-lacatestandardstocksolutionshouldbedividedintosingleusealiquotsandstoredat-20oC.

 

3.PrepareserialdilutionsofD-lactatestandard(0to1mM):

3.1   Add10μLofD-lactatestocksolution(fromStep2)into990µL1xPBSbuffertogenerate1mMD-lactatestandardsolution.

Note:DilutedD-lactatestandardsolutionisunstable,andshouldbeusedwithin4hours.

 

3.2   Take200μLof1mMD-lactatestandardsolutiontoperform1:3serialdilutionstoget300,100,30,10,3,1and0μMserialdilutionsofD-lactatestandard.

 

3.3   AddserialdilutionsofD-lactatestandardandD-lactatecontainingtestsamplesintoasolidblack96-wellmicroplateasdescribedinTables1and2.

 

Table1LayoutofD-lactatestandardsandtestsamplesinasolidblack96-wellmicroplate

BL

BL

TS

TS

….

….

 

 

 

 

 

 

Lac1

Lac1

….

….

….

….

 

 

 

 

 

 

Lac2

Lac2

 

 

 

 

 

 

 

 

 

 

Lac3

Lac3

 

 

 

 

 

 

 

 

 

 

Lac4

Lac4

 

 

 

 

 

 

 

 

 

 

Lac5

Lac5

 

 

 

 

 

 

 

 

 

 

Lac6

Lac6

 

 

 

 

 

 

 

 

 

 

Lac7

Lac7

 

 

 

 

 

 

 

 

 

 

               Note:Lac=D-LactateStandards,BL=BlankControl,TS=TestSamples.

 

Table2Reagentcompositionforeachwell

D-LactateStandard

BlankControl

TestSample

SerialDilutions*:50μL

DilutionBuffer:50μL

50μL

*Note:AddtheseriallydilutedD-Lactatestandardsfrom1μMto1mMintowellsfromLac1toLac7induplicate.

 

4.PrepareD-lactateassaymixture:

4.1   Add5mLofAssayBuffer(ComponentB)intoonebottleofEnzymeMix(ComponentA).

 

4.2   Add50µLNADstocksolution(100X,fromStep1)intothebottleofComponentA(fromStep4.1),and

mixwell.

Note:ThisD-lactateassaymixtureisenoughforone96-wellplate.TheunusedD-lactateassaymixtureshouldbedividedintosingleusealiquotsandstoredat-20oC.

 

5.RunD-lactateassay:

5.1   Add50μLofD-lactateassaymixture(fromStep4.2)toeachwellofD-lactatestandard,blankcontrol,andtestsamples(seeStep3.3)tomakethetotalD-lactateassayvolumeof100µL/well.

Note:Fora384-wellplate,add25μLofsampleand25μLofD-lactateassaymixtureintoeachwell.

 

5.2   Incubatethereactionatroomtemperaturefor30minutesto2hours,protectedfromlight.

 

5.3   MonitorthefluorescenceincreasewithafluorescenceplatereaderatEx/Em=530-570/590-600nm(optimalEx/Em=540/590nm,cutoffat570nm).

References&Citations
CitationExplorer

Arenobufagininhibitsprostatecancerepithelial-mesenchymaltransitionandmetastasisbydown-regulatingβ-catenin
Authors:LipingChen,WeiqianMai,MinfengChen,JianyangHu,ZhenjianZhuo,XuepingLei,LijuanDeng,JunshanLiu,NanYao,MaohuaHuang
Journal:PharmacologicalResearch(2017)

FibroblastActivationProteinAlpha-ActivatedTripeptideBufADIenolideAnti-tumorProdrugwithReducedCardiotoxicity
Authors:Li-JuanDeng,Long-HaiWang,Cheng-KangPeng,Yi-BinLi,Mao-HuaHuang,Min-FengChen,Xue-PingLei,MingQi,YunCen,Wen-CaiYe
Journal:JournalofMedicinalChemistry(2017)

NeuronalCultureMicroenvironmentsDeterminePreferencesinBioenergeticPathwayUse
Authors:JulianeSünwoldt,BertBosche,AndreasMeisel,PhilippMergenthaler
Journal:FrontiersinMolecularNeuroscience(2017):305

TheuseofKnockOutserumreplacement(KSR)inthreedimentionalrattesticularcellsco-culturemodel:Animprovedmalereproductivetoxicitytestingsystem
Authors:XiaofangZhang,LeiWang,XiaodongZhang,LijunRen,WenjingShi,YijunTian,JiangboZhu,TianbaoZhang
Journal:FoodandChemicalToxicology(2017)

Highglucoseandinterleukin1β-inducedapoptosisinhumanumbilicalveinendothelialcellsinvolvesindown-regulationofmonocarboxylatetransporter4
Authors:DongWang,QingjieWang,GaoliangYan,YongQiao,LingSun,BoqianZhu,ChengchunTang,YuchunGu
Journal:Biochemicalandbiophysicalresearchcommunications(2015):607--614

IntracellularATPdecreasemediatesNLRP3inflammasomeactivationuponnigericinandcrystalstimulation
Authors:JohjiNomura,AlexanderSo,MizuhoTamura,NathalieBusso
Journal:TheJournalofImmunology(2015):5718--5724

地衣芽胞杆菌对实验性结肠炎相关肝脏病变的疗效及机制影响
Authors:
Journal:世界华人消化杂志(2015):4399--4405


品牌介绍

AAT Bioquest常用产品报价单

产品名货号公司分类商城分类美金价
AAT Bioquest/Fluorescein, disodium salt *CAS 518-47-8*/2/100 mg2iFluor Rapid Tests酸碱缓冲液1088.00
AAT Bioquest/14-3-3 ζ (Ab-58) Antibody/8B0001/50 ug8B0001Signaling Intermediate Ab重组抗体2828.00
AAT Bioquest/Opioid Receptor (Phospho-Ser375) Antibody/8A0022/50 ug8A0022Phospho-Specific Ab受体2828.00
AAT Bioquest/iFluor™ 350 goat anti-mouse IgG (H+L)/16440/200 ug16440Fluorescent Anti-IgGs荧光染料1088.00
AAT Bioquest/trFluor™ Eu goat anti-mouse IgG (H+L)/16518/100 ug16518Fluorescent Anti-IgGs抗小鼠4278.00
AAT Bioquest/RPE-streptavidin conjugate/16900/100 ug16900Streptavidin Conjugates其他生物染料1378.00
AAT Bioquest/GCNT3 Antibody/8C14708/50 ug8C14708Signaling Intermediate Ab功能性抗体2828.00
AAT Bioquest/Amplite™ Luciferase Reporter Gene Assay Kit *Bright Glow*/12518/1 plate12518Reporter Gene Enzymes其它检测试剂盒2103.00
AAT Bioquest/ReadiLink™ Rapid mFluor™ Violet 450 Antibody Labeling Kit *Microscale Optimized for Labeling 50 &mi1100mFluor™ Dyes and Kits标记试剂盒2103.00
AAT Bioquest/Amplite™ Colorimetric Urea Quantitation Kit *Blue Color*/10058/200 Tests10058Diagnostic Molecules定量试剂盒2828.00
AAT Bioquest/Screen Quest™ CHO-Gqi Chimera Cell line/38101/Each38101cAMP GPCR Assays常规细胞株0.00
自营商城图标
厂家直采
全球直采 正品优价
正品保障图标
正品保障
厂家直发 有线跟踪
解放采购图标
正规清关
CIF100%正规报关,提供发票
及时交付图标
及时交付
限时必达 不达必赔