4000-520-616
欢迎来到免疫在线!(蚂蚁淘生物旗下平台)  请登录 |  免费注册 |  询价篮
aatbio(优势品牌)
主营:主营:研究并生产荧光和发光探针,信号转导研究的试剂
咨询热线电话
4000-520-616
当前位置: 首页 > 产品中心 > Other_kits > AAT Bioquest/Amplite™荧光葡萄糖-6-磷酸脱氢酶(G6PD)分析试剂盒/13806/200试验
商品详细AAT Bioquest/Amplite™荧光葡萄糖-6-磷酸脱氢酶(G6PD)分析试剂盒/13806/200试验
AAT Bioquest/Amplite™荧光葡萄糖-6-磷酸脱氢酶(G6PD)分析试剂盒/13806/200试验
AAT Bioquest/Amplite™荧光葡萄糖-6-磷酸脱氢酶(G6PD)分析试剂盒/13806/200试验
商品编号: 13806
品牌: aatbio
市场价: ¥85560.00
美元价: 51336.00
产地: 美国(厂家直采)
公司:
产品分类: 其它检测试剂盒
公司分类: Other_kits
联系Q Q: 3392242852
电话号码: 4000-520-616
电子邮箱: info@ebiomall.com
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
Glucose-6-phosphatedehydrogenase(G6PD)catalyzestheconversionofglucose-6-phosphateto6-phosphoglucono-?-lactone,thefirstandrate-limitingstepinthepentosephosphatepathway.Itiscriticalmetabolicpathwaythatsuppliesreducingenergytocells(suchaserythrocytes)bymaintainingthelevelofco-enzymenicotinamideadeninedinucleotidephosphate(NADPH),andfortheproductionofpentosesugars.TheproductionofNADPHisofgreatimportancefortissuesactivelyengagedinbiosynthesisoffattyacidsand/orisoprenoids,suchastheliver,mammaryglands,ADIposetissue,andtheadrenalglands.TheNADPHalsomaintainsthelevelofglutathioneinthesecellsthathelpsprotecttheredbloodcellsagainstoxidativedamage.DeficienciesinG6PDpredisposeindividualstonon-immunehemolyticanemia.AATBioquest"sAmplite™FluorimetricGlucose-6-PhosphateDehydrogenaseAssayKitprovidesasimple,sensitiveandrapidfluorescence-basedmethodfordetectingG6PDinbiologicalsamplessuchasserum,plasma,urine,aswellasincellculturesamples.Intheenzymecoupledassay,G6PDactivityisproportionallyrelatedtotheconcentrationofNADPHthatisspecificallymonitoredbyafluorogenicNADPHsensortoyieldahighlyredfluorescenceproduct.ThefluorescencesignalcanbereadwithafluorescencemicroplatereaderatEx/Em=540nm/590nm.WiththeG6PDassaykit,wewereabletodetectaslittleas1mU/mlG6PDina100µLreactionvolume.

Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.

1.PrepareNADPstocksolution(100X):

Add100µLofH2OintothevialofNADP(ComponentC)tomake100XNADPstocksolution.

 

2.PrepareG6PDstocksolution:

Add100µLofH2Oor1xPBSbufferintothevialofG6PDStandard(ComponentD)tomake100U/mLG6PDstandardsolution.

Note:TheunusedG6PDstandardstocksolutionshouldbedividedintosingleusealiquotsandstoredat-20oC.

 

3.PrepareserialdilutionsofG6PDstandard(0to300mU/mL):

3.1   Add10μLofG6PDstocksolution(fromStep2)into990µL1xPBSbuffertogenerate1000mU/mlG6PDstandardsolution.

Note:DilutedG6PDstandardsolutionisunstable,andshouldbeusedwithin4hours.

 

3.2   Take15μLof1000mU/mlG6PDstandardsolution(fromstep3.1)into485uLof1xPBSbuffertogenerate30mU/mLG6PD,andthenperform1:3serialdilutionstoget10,3.3,1.1,0.37,0.12,0.04and0mU/mLserialdilutionsofG6PDstandard.

 

3.3   AddserialdilutionsofG6PDstandardandG6PDcontainingtestsamplesintoasolidblack96-wellmicroplateasdescribedinTables1and2.

 

Table1:LayoutofG6PDstandardsandtestsamplesinasolidblack96-wellmicroplate

BL

BL

TS

TS

….

….

 

 

 

 

 

 

G6PD1

G6PD1

….

….

….

….

 

 

 

 

 

 

G6PD2

G6PD2

 

 

 

 

 

 

 

 

 

 

G6PD3

G6PD3

 

 

 

 

 

 

 

 

 

 

G6PD4

G6PD4

 

 

 

 

 

 

 

 

 

 

G6PD5

G6PD5

 

 

 

 

 

 

 

 

 

 

G6PD6

G6PD6

 

 

 

 

 

 

 

 

 

 

G6PD7

G6PD7

 

 

 

 

 

 

 

 

 

 

               Note:G6PD=D-Glucose-6-PhosphateDehydrogenaseStandards,BL=BlankControl,TS=TestSamples.

 

Table2Reagentcompositionforeachwell

G6PDStandard

BlankControl

TestSample

SerialDilutions*:50μL

DilutionBuffer(PBS):50μL

50μL

*Note:AddtheseriallydilutedG6PDstandardsfrom0.04mU/mLto30mU/mLintowellsfromG6PD1toG6PD7induplicate.

 

4.PrepareG6PDassaymixture:

4.1   Add5mLofAssayBuffer(ComponentB)intoonebottleofEnzymeProbe(ComponentA).

 

4.2   Add50µLNADPstocksolution(100X)intothebottleofComponentA (fromStep4.1),andmixwell.

Note:ThisG6PDassaymixtureisenoughforone96-wellplate.Theunusedassaymixtureshouldbedividedintosingleusealiquotsandstoredat-20oC.

 

5.RunG6PDassay:

5.1   Add50μLofG6PDassaymixture(fromStep4.2)toeachwellofG6PDstandard,blankcontrol,andtestsamples(seeStep3.3)tomakethetotalassayvolumeof100µL/well.

Note:Fora384-wellplate,add25μLofsampleand25μLassaymixtureintoeachwell.

 

5.2   Incubatethereactionatroomtemperaturefor30minutesto2hours,protectedfromlight.

 

5.3   MonitorthefluorescenceincreasewithafluorescenceplatereaderatEx/Em=540/590nm,cutoffat570nm.

References&Citations
PrinterFriendlyVersion

1.   LegezaB,BalazsZ,NashevLG,OdermattA.(2013)Themicrosomalenzyme17beta-hydroxysteroiddehydrogenase3facesthecytoplasmandusesNADPHgeneratedbyglucose-6-phosphatedehydrogenase.Endocrinology,154,205.

2.   HeckerPA,MapangaRF,KimarCP,RibeiroRF,Jr.,BrownBH,O"ConnellKA,CoxJW,ShekarKC,AsemuG,EssopMF,StanleyWC.(2012)Effectsofglucose-6-phosphatedehydrogenasedeficiencyonthemetabolicandcardiacresponsestoobesogenicorhigh-fructosediets.AmJPhysiolEndocrinolMetab,303,E959.

3.   Al-MusawiBM,Al-AllawiN,ABDul-MajeedBA,EissaAA,JubraelJM,HamamyH.(2012)Molecularcharacterizationofglucose-6-phosphatedehydrogenasedeficientvariantsinBaghdadcity-Iraq.BMCBloodDisord,12,4.

4.   ShahSS,DiakiteSA,TraoreK,DiakiteM,KwiatkowskiDP,RockettKA,WellemsTE,FairhurstRM.(2012)Anovelcytofluorometricassayforthedetectionandquantificationofglucose-6-phosphatedehydrogenasedeficiency.SciRep,2,299.

5.   OrimadegunAE,SodeindeO.(2011)Glucose-6-phosphatedehydrogenasestatusandseverityofmalarialanaemiainNigerianchildren.JInfectDevCtries,5,792.

6.   SchneiderAM,RawatD,WeinsteinLS,GupteSA,RichardsWO.(2012)EffectsoflaparoscopicRoux-en-Ygastricbypassonglucose-6phosphatedehydrogenaseactivityinobesetype2diabetics.SurgEndosc,26,823.

7.   MillimonoTS,LouaKM,RathSL,RelvasL,BentoC,DiakiteM,JarvisM,DariesN,RibeiroLM,MancoL,KaedaJS.(2012)Highprevalenceofhemoglobindisordersandglucose-6-phosphatedehydrogenase(G6PD)deficiencyintheRepublicofGuinea(WestAfrica).Hemoglobin,36,25.

8.   GavriliukLA,KorchmaruIF,RobuMV,LysyiLT.(2011)[Glutathione-dependentenzymesandglucose-6-phosphatedehydrogenaseofbloodinpatientswithlymphosarcoma(non-Hodgkin"sdisease)].BiomedKhim,57,225.

9.   CarterN,PambaA,DuparcS,WaitumbiJN.(2011)Frequencyofglucose-6-phosphatedehydrogenasedeficiencyinmalariapatientsfromsixAfricancountriesenrolledintworandomizedanti-malarialclinicaltrials.MalarJ,10,241.

10.   EandiEberleS,GarciaRosolenN,UrtasunC,SciuccatiG,DiazL,SaviettoV,CandasA,AvalosGomezV,CervioC,BonduelM,FeliuTorresA.(2011)[Glucose6phosphatedehydrogenasedeficiency:acaseseries].ArchArgentPediatr,109,354.


ViewAllAsPDF
品牌介绍
美国AAT Bioquest Inc.(前身是ABD Bioquest,Inc.)是一家为从事生命科学研究、诊断研发及药物开发的科学家研发、生产和销售生物分析研究试剂和试剂盒的公司。公司致力于光谱学检测领域,包括吸收(颜色)、荧光和发光技术。AAT Bioquest的产品帮助全世界的科学家和生物医药研究者更好的了解生物化学、免疫学、细胞生物学和分子生物学等领域。AAT Bioquest会不断研发新产品,快速地丰富各个领域的产品。 1)提供反应探针和发光探针,生物素和端粒酶能够应用于标记药物小分子和生物聚合物,如蛋白、核酸以及其他碳水化合物; 2)研究并生产荧光和发光探针用于检测蛋白,核酸和活细胞; 3)不断推出新型的荧光和发光探针用于检测多种酶,特别是检测水解酶和氧化还原酶类; 4)致力于开发用于信号转导研究的试剂; 5)提供生理和神经探针,特别是钙离子指示剂和膜电位探针。
品牌分类
淋巴细胞信号传导 GPCR抗体 羰基活性生物素 细胞代谢 胺反应生物素 链霉亲和素结合物 生物素化抗IgGs 乙酰基特异性抗体 细胞骨架抗体
联络我们
服务热线:4000-520-616
(限工作日9:00-18:00)
QQ :1570468124
手机:18915418616