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:
- Specificallymeasureepithelialapoptosisincellcultureapplications
- Theassayhasadynamicrangeandsensitivitysuitableforinvitrowork
- SpecificquantificationmeasurementtoolforapoptosisinCK18positivecells
- Measuresastable,accumulatedproductallowingformeasurementat1latertimepoint
- SandwichELISAina96wellplateinaconvenientready-to-useformat
- Easytoperform,onlyaminimumofpipettingstepsrequired
- CK18ispresentinsimple epithelialcellsonly,thusincreasingspecificityofmeasurementinepithelial-basedmodels
- Powerfultoolforinvitrocharacterizationofapoptosis-inducingdrugs.
- Mesauresformationofacapase-cleavageproductintheinteriorofspheroids
- Useoftissueslicesfromexvivotumorstomeasureapoptosisofcandidatedrugs
- Keyadvantageinco‐culturesystemstodeterminedrugeffectonepithelialtissue
CK18KitComposition:
Reagents,Packaging,Storage,andStABIlity
- M6CoatedMicrostrips: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).
M65andM30kitsrobustlydetectcelltoxicityandapoptosisinHepaRGcells invitroexperiments. Aadditionalkeyfeatureisthatthe CK18kits measurean accumulated caspase-derivedproduct,sothereisnorisktomisstimeofapoptosisandtransientcaspaseactivation(Seefigurebelow).
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2.NaHCO3,Ba(OH)3,H2SO4
3.HCL,NaAlO2,NaHSO4
4.Ca(OH)2,Na2CO3,BaCO3
谢谢了
要原因
请问有什么方法检测细胞内活性氧更好,各位有什么经验,及相关试剂盒推荐一下。
A.H2SO4Na2SO4AgNO3BaCl2.
B.NaOHNa2CO3NaHSO4MgCl2
C.CaCl2NaNO3MgSO4BaCl2
D.HNO3KOHKClK2CO3
请问下有无同学需要H37RA的?我是做EAE模型的,上个月购买了BDDifco公司的H37RA(货号),因为购买的时候只能整盒6支购买,但我们用不了那么多,所以想问问有无同学需要的,100mg/支,800元/支或用等价试剂交换。地址广州。有需要的请私信,谢谢!
(1)优级纯试剂 亦称保证试剂,为一级品,纯度高,杂质极少,主要用于精密分析和科学研究,常以GR表示。
(2)分析纯试剂 亦称分析试剂,为二级品,纯度略低于优级纯,杂质含量略高于优级纯,适用于重要分析和一般性研究工作,常以AR表示。
(3)化学纯试剂 为三级品,纯度较分析纯差,但高于实验试剂,适用于工厂、学校一般性的分析工作,常以CP表示。
(4)实验试剂 为四级品,纯度比化学纯差,但比工业品纯度高,主要用于一般化学实验,不能用于分析工作,常以 LR表示。
以上按试剂纯度的分类法已在我国通用。根据化学工业部颁布的“化学试剂包装及标志”的规定,化学试剂的不同等级分别用各种不同的颜色来标志,见表1。
表1 我国化学试剂的等级及标志
bhclna2so4nano3na2co3
chclnaohna2co3nacl
dba(oh)2nahco3alcl3nahso4
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