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™ Colorimetric Acetylcholinesterase Assay Kit/11400/200 Tests
产品编号:11400
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 Acetylcholinesterase Assay Kit/11400/200 Tests
商品介绍
Overview
PrinterFriendlyVersion

Ex/Em(nm)410/None
MWN/A
CAS#N/A
SolventN/A
StorageF/D/L
CategoryNeuroBIOLOGy
NeurologicalEnzymes
RelatedHydrolyticEnzymes
MicroplateReaders
Acetylcholinesterase,alsoknownasAChE,isanenzymethatdegrades(throughitshydrolyticactivity)theneurotransmitteracetylcholine,producingcholineandanacetategroup.Itismainlyfoundatneuromuscularjunctionsandcholinergicsynapsesinthecentralnervoussystem,whereitsactivityservestoterminatesynaptictransmission.AChEhasaveryhighcatalyticactivity-eachmoleculeofAChEdegradesabout5000moleculesofacetylcholinepersecond.Acetylcholinesteraseisalsofoundontheredbloodcellmembranes,whereitconstitutestheYtbloodgroupantigen.Acetylcholinesteraseexistsinmultiplemolecularforms,whichpossesssimilarcatalyticproperties,butdifferintheiroligomericassemblyandmodeofattachmenttothecellsurface.ThisAmplite™ColorimetricAcetylcholinesteraseAssayKitprovidesaconvenientmethodforthedetectingAChEactivity.ThekitusesDTNBtoquantifythethiolcholineproducedfromthehydrolysisofacetylthiolcholinebyAChE.TheabsoprtionintensityofDTNBadductisproportionaltotheformationofthiolcholine,thustheAChEactivity.

Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.

1.Preparestocksolutions:

1.1   20XDTNBstocksolution:Add0.6mLofAssayBuffer(ComponentB)intothevialofDTNB(ComponentA)tomake20XDTNBstocksolution.

Note:TheunusedDTNBstocksolutionshouldbedividedintosingleusealiquots.Storeat-20oCandkeepfromlight.

 

1.2   20XAcetylthiocholinestocksolution:Add0.6mLofddH2Ointothevialofacetylthiocholine(Component C).

Note:Theunused20Xacetylthiocholinestocksolutionshouldbedividedintosingleusealiquotsandstoredat‑20oC.

1.3   Acetylcholinesterasestocksolution:Add100µLofddH2Owith0.1%BSAintothevialofacetylcholinesterasestandard(ComponentD)tomakea50units/mLacetylcholinesterasestocksolution.

Note:Theunusedacetylcholinesterasestocksolutionshouldbedividedintosingleusealiquotsandstoredat-20oC.

 

2.Prepareacetylthiocholinereactionmixture:

PreparetheacetylthiocholinereactionmixtureaccordingtoTable1andkeepfromlight.

 

Table1Acetylthiocholinereactionmixtureforone96-wellplate

Components

Volume

AssayBuffer(ComponentB)

4.5mL

20XDTNBStockSolution(fromStep1.1)

250µL

20XAcetylthiocholineStocksolution(fromStep1.2)

250µL

Totalvolume

5mL

 

3.Prepareserialdilutionsofacetylcholinesterasestandard(0to1000mU/mL):

3.1   Add20μLof50units/mLacetylcholinesterasestocksolution(fromStep1.3)to980µLofassaybuffer(ComponentB)togenerate1000mU/mLacetylcholinesterasestandardsolution.

Note:Dilutedacetylcholinesterasestandardsolutionisunstableandshouldbeusedwithin4hours.

 

3.2   Take200μLof1000mU/mLacetylcholinesterasestandardtoperform1:3serialdilutionstoget300,100,30,10,3,1and0mU/mLserialdilutionsofacetylcholinesterasestandard.

 

3.3   Addserialdilutionsofacetylcholinesterasestandardandacetylcholinesterase-containingtestsamplesintoawhite/clearbottom96-wellmicroplateasdescribedinTables2and3.

Note:Treatcellsortissuesamplesasdesired.

 

Table2.Layoutofacetylcholinesterasestandardsandtestsamplesinawhite/clearbottom96-wellmicroplate

BL

BL

TS

TS

….

….

 

 

 

 

 

 

AS1

AS1

….

….

….

….

 

 

 

 

 

 

AS2

AS2

 

 

 

 

 

 

 

 

 

 

AS3

AS3

 

 

 

 

 

 

 

 

 

 

AS4

AS4

 

 

 

 

 

 

 

 

 

 

AS5

AS5

 

 

 

 

 

 

 

 

 

 

AS6

AS6

 

 

 

 

 

 

 

 

 

 

AS7

AS7

 

 

 

 

 

 

 

 

 

 

Note:AS=AcetylcholinesteraseStandards;BL=BlankControl;TS=TestSamples

 

Table3.Reagentcompositionforeachwell

AcetylcholinesteraseStandard

BlankControl

TestSample

SerialDilutions*:50μL

AssayBuffer:50μL

50μL

*Note:Addtheserialdilutionsofacetylcholinesterasestandardfrom1to1000mU/mLintowellsfromAS1toAS7induplicate.

 

4.Runacetylcholinesteraseassay:

4.1   Add50μLofacetylthiocholinereactionmixture(fromStep2.1)toeachwelloftheacetylcholinesterasestandard,blankcontrol,andtestsamples(seeStep3.3)tomakethetotalacetylcholinesteraseassayvolumeof100µL/well.

Note:Fora384-wellplate,add25μLofsampleand25μLofacetylthiocholinereactionmixtureineachwell.

4.2   Incubatethereactionfor10to30minutesatroomtemperature,protectedfromlight.

4.3   Monitortheabsorbanceincreasewithanabsorbancemicroplatereaderat410±5nm.

References&Citations
CitationExplorer

HepaticvagusnerveregulatesKupffercellactivationviaα7nicotinicacetylcholinereceptorinnonalcoholicsteatohepatitis
Authors:TakahiroNishio,KojiroTaura,KeikoIwaisako,YukinoriKoyama,KazutakaTanabe,GenYamamoto,YukihiroOkuda,YoshinobuIkeno,KenjiYoshino,YosukeKasai
Journal:JournalofGastroenterology(2017):1--12

ProtectiveEffectofα-LipoicAcidagainstα-Cypermethrin-InducedChangesinRatCerebellum
Authors:HElsawy,MAAl-Omair,ASedky,LAl-Otaibi
Journal:JournalofChemicalNeuroanatomy(2017)

SpirulinamaximaExtractAmelioratesLearningandMemoryImpairmentsviaInhibitingGSK-3βPhosphorylationInducedbyIntracerebroventricularInjectionofAmyloid-β1--42inMice
Authors:Eun-JeongKoh,Kui-JinKim,Ji-HyeonSong,JiaChoi,HyeonYongLee,Do-HyungKang,HoJinHeo,Boo-YongLee
Journal:InternationalJournalofMolecularSciences(2017):2401

Anetholerestoresdelayedgastricemptyingandimpairedgastricaccommodationinrodents
Authors:TeitaAsano,ShujiAida,ShintaroSuemasu,TohruMizushima
Journal:Biochemicalandbiophysicalresearchcommunications(2016):125--130

DietaryanddonepezilmodulationofmTORsignalingandneuroinflammationinthebrain
Authors:KalavathiDasuri,LeZhang,SunOKFernandezKim,AnnadoraJBruce-Keller,JeffreyNKeller
Journal:BiochimicaetBiophysicaActa(BBA)-MolecularBasisofDisease(2016):274--283

DissolvedorganicmatterorsaltschangethebioavailABIlityprocessesandtoxicityofthenanoscaletetravalentleadcorrosionproductPbO2tomedakafish
Authors:Chun-WeiChiang,Ding-QuanNg,Yi-PinLin,Pei-JenChen
Journal:EnvironmentalScience&Technology(2016):11292--11301

FunctionaliseddihydroazopyrimidinederivativesfromMorita--Baylis--Hillmanacetates:synthesisandstudiesagainstacetylcholinesteraseasitsinhibitors
Authors:EedaKotiReddy,CRemya,AyyiliathMSajith,KVDileep,CSadasivan,ShaikAnwar
Journal:RSCAdvances(2016):77431--77439

Gongjin-DanEnhancesHippocampalMemoryinaMouseModelofScopolamine-InducedAmnesia
Authors:Jin-SeokLee,Sung-ShinHong,Hyeong-GeugKim,Hye-WonLee,Won-YongKim,Sam-KeunLee,Chang-GueSon
Journal:PloSone(2016):e0159823

VisceralADIposetissue-derivedserineproteaseinhibitoraugmentsacetylcholine-inducedrelaxationviatheinhibitionofacetylcholineesteraseactivityinratisolatedmesentericartery
Authors:SKameshima,KYamada,TMorita,MOkada,HYamawaki
Journal:ActaPhysiologica(2016):203--210

Z-GuggulsteroneImprovestheScopolamine-InducedMemoryImpairmentsThroughEnhancementoftheBDNFSignalinC57BL/6JMice
Authors:ZhuoChen,ChaoHuang,WenbinDing
Journal:NeuRochemicalResearch(2016):3322--3332


ViewMoreCitations
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