产品说明
Description:TheM30CytoDeath™ELISAisanovelresearchkit developedforcellcultureapplications.TheM30CytoDeath™assayhasadynamicrangeandsensitivitysuitableforinvitrowork.M30CytoDeath™ELISAisapowerfuldrugscreeningtool.Itisusefulforinvitrocharacterizationofapoptosis-inducingdrugs,includingestablishmentoftimecoursekineticsanddoseresponserelationships.TheeffectsoflargenumbersofsiRNAsonapoptosiscanbetestedinthe96wellformat.Theassayoffersauniquepossibilitytoquantifyapoptosisofepitheliallyderivedcellsinmulticellularspheroidsandorganculturesystems.TheM30CytoDeath™ELISAisbasedontheM30antibody(detectingtheAsp396caspase-cleavagesiteonCK18)andantibodyM6.ThiscombinationisdifferentfromthatusedintheM30-Apoptosense®ELISA.TheM30CytoDeath™ELISAisnotsuitableforserum/plasma.CK18KitAdvantages:SpecificallymeasureepithelialapoptosisincellcultureapplicationsTheassayhasadynamicrangeandsensitivitysuitableforinvitroworkSpecificquantificationmeasurementtoolforapoptosisinCK18positivecellsMeasuresastable,accumulatedproductallowingformeasurementat1latertimepointSandwichELISAina96wellplateinaconvenientready-to-useformatEasytoperform,onlyaminimumofpipettingstepsrequiredCK18ispresentinsimple epithelialcellsonly,thusincreasingspecificityofmeasurementinepithelial-basedmodelsPowerfultoolforinvitrocharacterizationofapoptosis-inducingdrugs.Mesauresformationofacapase-cleavageproductintheinteriorofspheroidsUseoftissueslicesfromexvivotumorstomeasureapoptosisofcandidatedrugsKeyadvantageinco‐culturesystemstodeterminedrugeffectonepithelialtissueCK18KitComposition:Reagents,Packaging,Storage,andStABIlityM6CoatedMicrostrips:Onemicroplate,12stripswith8wellseach,96dry wellsintotal.ThewellsarecoatedwithmousemonoclonalK18antibodyM6. Themicroplateissealedinanaluminiumbag,whichcontainsadesiccatingdevice. Ifnotallthestripsareused,resealthebagandkeepthedesiccatingdevice inside.Readyforuse!M30CytoDeathHRPConjugate:Concentrate(24×conc.).Onevialcontaining 0.4mLofmousemonoclonalM30antibody(anti-K18Asp396neo-epitope)conjugated withhorserADIshperoxidase(HRP)inaphosphatebufferwithprotein stabilizers.Preservativeadded.ShouldbedilutedwithM30CytoDeathConjugate DilutionBuffer(seeInstructionsforUsesection“ComponentPreparation”onpage9).Note!Do notexposetolight!M30CytoDeathConjugateDilutionBuffer:Onevialcontaining11mLofphosphate bufferwithproteinstabilizersfordilutionoftheM30CytoDeathHRPConjugate. Preservativeadded.Greencolored.M30CytoDeathStandards:StandardZerocontaining0.5mLofphosphate bufferwithfetalcalfserum(FCS).StandardsLow,MediumandHigh,0.5mL each,containingstandardmaterialinaphosphatebufferwithFCS.Thevaluesof theStandardsare0U/L(Zero),250U/L(Low),1000U/L(Medium)and3000U/L (High).Preservativeadded.Yellowcolored.Readyforuse!StandardZerocan beusedfordilutionsofsamples>3000U/L.WashTablet:Onetabletfor500mLofpreparedwashsolution.Dissolvethe WashTabletin500mLoffreshdeionisedwater.TMBSubstrate:Onebottlecontaining22mLofTMB(3,3’,5,5’-Tetramethylbenzidine) Substrate.Note!Donotexposetolight!Readyforuse!StopSolution:Onevialcontaining7mLof1.0Msulphuricacid.Readyforuse!SealingTape:One(1)sheet. MeasurementPrinciple:TheM30CytoDeath™ELISAisasolid-phasesandwichenzymeimmunoassay. StandardsandsamplesreactwithasolidphasecaptureantibodyM6directed againstK18andtheHRP(horseradishperoxidase)conjugatedM30antibody directedagainsttheK18Asp396neo-epitope.Unboundconjugateisremoved byawashingstep.TMBSubstrateisadded.Thecolordevelopmentisstoppedandtheabsorbanceisread.Theresultingcolorisdirectlyproportionaltothe concentrationoftheanalyte. Byplottingastandardcurvefromknownconcentrationsversusmeasuredabsorbance, theamountofantigeninthesamplecanbecalculated.Theconcentration oftheantigenisexpressedasunitsperliter(U/L).Background:Inastudy presentedattheSOTMeetingin2014,toxicitytestingoncryopreserveddifferentiatedHepaRG™cellswasperformedusingCK18kits.HepaRG™cellsexhibitmanycharacteristicsofprimaryhumanhepatocytes,includingmorphologyandexpressionofkeymetabolicenzymes,nuclearreceptors,anddrugtransporters.UnlikeHepG2cells,HepaRG™cellshavehighP450activityandcompleteexpressionofallnuclearreceptors.HepaRG™wasexposedtodifferentcompounds,includingparacetamol,chloropromazine,rotenone,rosiglitazoneandomeprazole.ToxicityandapoptosiswereassayedfromcollectedcellculturesupernatantsmeasuringM65andM30levels.M65resultswereconsistentwithcellviability,whereas,M30resultsindicatedthatapoptosiswasinducedatlowerdrugconcentrationswhilenecrosiswasmoreprominentathigherones(seefigurebelow).HumanHepRG™hepatocytestreatedwithdifferentcompounds.Toxicityandapoptosiswereassessedfromcollectedsupernatantsmeasuringfull-lengthK18(M65)andcaspase-cleavedK18(ccK18,M30)usingM65EpiDeath®andM30CytoDeath™ELISA.M65andM30kitsrobustlydetectcelltoxicityandapoptosisinHepaRGcells invitroexperiments. Aadditionalkeyfeatureisthatthe CK18kits measurean accumulated caspase-derivedproduct,sothereisnorisktomisstimeofapoptosisandtransientcaspaseactivation(Seefigurebelow).
Diapharma使命宣言
位于俄亥俄州西切斯特的Difarma Group,Inc.在诊断和研究领域销售止血、血栓形成、血小板功能测试、仪器和凋亡产品,并提供强大的技术能力和经验,以确保满足或超过客户的期望。
地黄止血显色凝块酶联免疫吸附试验试剂盒
历史
1997年1月1日,由俄亥俄州富兰克林市的Pharmacia Hepar,Inc.成立,最初是Chromogenix基质和分析的独家美国和加拿大经销商。
四分之一个多世纪前,Chromogenix开发了第一个显色底物技术,其前身是Kabi Diagnostica。卡比后来与法玛西亚合并。希帕玛目前的一些员工在法玛西亚肝素制造厂的显色部门工作。
1998年,夏帕玛搬到了俄亥俄州的西切斯特,至今仍在那里。多年来,迪法玛扩大了其产品线,包括各种止血、细胞死亡、血小板功能、生态毒理学、化验、试剂、抗体和高级制造商的仪器。
2017年,夏帕玛庆祝了20年的成功
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