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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/Cell Meter™ Bacterial Viability Assay Kit/22400/200 Tests
产品编号:22400
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/Cell Meter™ Bacterial Viability Assay Kit/22400/200 Tests
商品介绍
Overview
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Ex/Em(nm)484/520&630
MWN/A
CAS#N/A
SolventDMSO
StorageF/D/L
CategoryMicroBIOLOGy
FlowCytometry
RelatedFluorescenceImaging
AATBioquest"sMycolight™BacterialViABIlityAssayKitprovidestwo-colorfluorescenceassayofbacterialviabilityinbothgram-positiveandnegativebacterialcell.ThekitutilizesthemixtureofourgreenfluorescentnucleicacidstainMycoLight™Greenandthered-fluorescentnucleicacidstainpropidiumiodide.Whenusedalone,theMycoLight™Greenstaingenerallylabelsallbacteria(liveanddead)inapopulation.Incontrast,propidiumiodidepenetratesonlybacteriawithdamagedmembranes,causingareductionintheMycoLight™Greenstainfluorescencewhenbothdyesarepresent.Thus,withanappropriatemixtureoftheMycoLight™Greenandpropidiumiodidestains,livebacteriawithintactcellmembranesemitsgreenfluorescence,whereasdeadordyingbacteriawithdamagedmembranesgivesredfluorescence.TheMycolight™BacterialViabilityAssayKitisarobusttoolformonitoringtheviabilityofbacterialpopulationsasafunctionofthemembraneintegrityofthecell.Stainedcellscanbemonitoredfluorimetricallyat510-530nm(FITCfilter)and600-660nm(Texasredfilter)withexcitationat488nm,themostcommonexcitationlightsource.
SpectrumAdvancedSpectrumViewer

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Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.

Note1:Thawkitcomponentsatroomtemperatureandcentrifugebrieflybeforestartingyourexperiment.Note2:TheKithasbeentestedatlogarithmicallygrowingculturesofthefollowingbacterialspecies:Bacilluscereus,B.subtilis,Clostridiumperfringens,Escherichiacoli,Klebsiellapneumoniae,Micrococcusluteus,Mycobacteriumphlei,Pseudomonasaeruginosa,P.syringae,Salmonellaoranienburg,Serratiamarcescens,Shigellasonnei,StaphylococcusaureusandStreptococcuspyogenes.Agrobacteriumtumefaciens,Edwardsiellaictaluri,Eurioplasmaeurilytica,Lactobacillussp.,Mycoplasmahominus,Propionibacteriumsp.,ProteusmirabilisandZymomonassp.Note3:Thefollowingistherecommendedprotocolforbacterialstaining.Theprotocolonlyprovidesaguideline,shouldbemodifiedaccordingtothespecificneeds.

1.      Growbacteriainanyappropriatemedium.Bestresultsforhealthybacteriaareobtainedfromlog-phasecultures.Dilutethebacterialcultureto~106cto108cellspermLin0.85%NaClorappropriatebuffer.PreparesufficientsUSPensiontoprovide500µLpertestforflowcytometryor100µLpertestfor96-wellplate.

Note:Removetracesofgrowthmediumbeforestainingbacteria.Asinglewashstepisusuallysufficienttoremovesignificanttracesofinterferingmediacomponentsfromthebacterialsuspension.Phosphatewashbuffersarenotrecommendedbecausetheyappeartodecreasestainingefficiency.

2.      MixequalvolumeofMycoLight™Green(ComponentA)andpropidiumiodide(ComponentB)inamicrofugetubetohave250xstainingdyemixture.

3.      Add4µLofthe250xstainingdyemixture(fromstep2)toeachmLofthebacterialsuspension.Mixwellandincubateatroomtemperaturefor15minutes.Protectfromlight.

4.      Thestainedbacterialcellscanbeanalyzedbyafluorescencemicroscope,fluorescentmicroplatereaderorflowcytometry.

5.      Thefluorescencefrombothliveanddeadbacteriamaybeviewedsimultaneouslywithanystandardfluoresceinlongpassfilterset.Alternatively,thelive(greenfluorescent)anddead(redfluorescent)cellsmaybeviewedseparatelywithfluoresceinandTexasRedfiltersets.

References&Citations
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1.   Hu,W.;Murata,K.;Zhang,D.,ApplicabilityofLIVE/DEADBacLightstainwithglutaraldehydefixationforthemeasurementofbacterialabundanceandviabilityinrainwater.JEnvironSci(China)2017,51,202-213.

2.   Karkashan,A.;Khallaf,B.;Morris,J.;Thurbon,N.;Rouch,D.;Smith,S.R.;Deighton,M.,ComparisonofmethodologiesforenumeratinganddetectingtheviabilityofAscariseggsinsewagesludgebystandardincubation-microscopy,theBacLightLive/Deadviabilityassayandothervitaldyes.WaterRes2015,68,533-44.

3.   Wahman,D.G.;Schrantz,K.A.;Pressman,J.G.,DeterminationoftheeffectsofmediumcompositiononthemonochloraminedisinfectionkineticsofNitrosomonaseuropaeabythepropidiummonoazidequantitativePCRandLive/DeadBacLightmethods.ApplEnvironMicrobiol2010,76(24),8277-80.

4.   Wahman,D.G.;Wulfeck-Kleier,K.A.;Pressman,J.G.,MonochloraminedisinfectionkineticsofNitrosomonaseuropaeabypropidiummonoazidequantitativePCRandLive/deadBacLightmethods.ApplEnvironMicrobiol2009,75(17),5555-62.

5.   Berney,M.;Hammes,F.;Bosshard,F.;Weilenmann,H.U.;Egli,T.,AssessmentandinterpretationofbacterialviabilitybyusingtheLIVE/DEADBacLightKitincombinationwithflowcytometry.ApplEnvironMicrobiol2007,73(10),3283-90.

6.   Leuko,S.;Legat,A.;Fendrihan,S.;Stan-Lotter,H.,EvaluationoftheLIVE/DEADBacLightkitfordetectionofextremophilicarchaeaandvisualizationofmicroorganismsinenvironmentalhypersalinesamples.ApplEnvironMicrobiol2004,70(11),6884-6.

7.   Boulos,L.;Prevost,M.;Barbeau,B.;Coallier,J.;Desjardins,R.,LIVE/DEADBacLight:applicationofanewrapidstainingmethodfordirectenumerationofviableandtotalbacteriaindrinkingwater.JMicrobiolMethods1999,37(1),77-86.


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