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Cell Death Detection ELISA PLUS 细胞凋亡

CellDeathDetectionELISAPLUS，现货供应！Roche经典产品！

Application

TheCellDeathDetectionELISAPLUSphotometricenzymeimmunoassayisusedforthequantitativeinvitrodeterminationofcytoplasmichistone-associatedDNAfragments(mono-andoligonucleosomes)afterinducedcelldeath.
Thekitcontainsastopsolutionwhichallowsuserstoterminatethesubstratereactionandruntheassayunderdefinedconditions,makingitsuitableforuseinhigh-throughputapplications.

Benefits
  • One-stepELISA
  • Highsensitivity:(5x102cells/ml)
  • Fastperformance:(3-4hours)
  • Positivecontrolincluded
  • Noprelabelingofcellsnecessary
  • NonrADIoactiveassaysystem
  • Nospeciesrestriction
  • Easyhandling
  • Lowbackground
  • Suppressedhumananti-mousefactor
  • Function-tested
ProductDescription

Assaytime:
3-4hours
Negativecontrol:
Dependingoncellcultureconditions,eachexponentiallygrowingpermanentcellculturecontainsacertainamountofdeadcells(typicallyapproximately3-8%).Intheimmunoassay,theseinherentdeadcellsintheuntreatedsample(withoutacell-death-inducingagent)willcauseacertainabsorbancevalue(negativecontrol).
Positivecontrol:
ADNA-histonecomplexservesasapositivecontrol.
Samplematerial:
Cytoplasmicfractions(lysates)ofcelllines,cellsexvivo,cellculturesupernatants,andserumorplasma
Sensitivity:
Theexactdetectionlimitofdying/deadcellsinaparticularsamplestronglydependsonthekineticsofcelldeath,thecytotoxicagentused,andtheamountofaffectedcellsinthetotalcellpopulation.UsingU937/camptothecin(CAM)asacellularmodelsystemforcelldeath,theimmunoassayallowsthespecificdetectionofmono-andoligonucleosomesinthecytoplasmicfractionof125cellequivalents/well.
Specificity:
Anti-histonereactswiththehistonesH1,H2A,H2B,H3,andH4ofvariousspecies(e.g.,human,mouse,rat,hamster,cow,opossum,Xenopus).Anti-DNAbindstosingle-anddouble-strandedDNA.Therefore,theELISAallowsthedetectionofmono-andoligonucleosomesfromvariousspecies,andmaybeappliedtomeasureapoptoticcelldeathinmanydifferentcellsystems.

BackgroundInformation
Twodistinctformsofeukaryoticcelldeathcanbeclassifiedbymorphologicalandbiochemicalcriteria:necrosisandapoptosis.NecrosisisaccompaniedbyincreasedionpermeABIlityoftheplasmamembrane;thecellsswellandtheplasmamembraneruptureswithinminutes(osmoticlysis).Apoptosisischaracterizedbymembraneblebbing(zeiosis),condensationofcytoplasm,andtheactivationofanendogenousendonuclease.ThisCa2+-andMg2+-dependentnucleasecleavesdouble-strandedDNAatthemostaccessIBLeinternucleosomallinkerregion,generatingmono-andoligonucleosomes.Incontrast,theDNAofthenucleosomesistightlycomplexedwiththecorehistonesH2A,H2B,H3,andH4,andisthusprotectedfromcleavagebytheendonuclease.TheyieldedDNAfragmentsarediscretemultiplesofan180-bpsubunit,detectedasa“DNAladder”onagarosegelsafterextractionandseparationofthefragmentedDNA.Theenrichmentofmono-andoligonucleosomesinthecytoplasmoftheapoptoticcellisduetothefactthatDNAdegradationoccursseveralhoursbeforeplasmamembranebreakdown.
Contents
  1. Anti-histone-biotin(cloneH11-4)
  2. Anti-DNA-POD(cloneMCA-33)
  3. PositiveControl
  4. IncubationBuffer
  5. LysisBuffer
  6. SubstrateBuffer
  7. ABTSSubstrateTablet
  8. ABTSStopSolution
  9. Microplate(streptavidin-coated)
  10. AdhesiveCoverFoils
Principle

Theassayisbasedonthequantitative“sandwichenzymeimmunoassay”principleusingmousemonoclonalantibodiesdirectedagainstDNAandhistones.Thisallowsthespecificdeterminationofmono-andoligonucleosomesinthecytoplasmicfractionofcelllysates.Thesamplesareplacedintoastreptavidin-coatedmicroplateandincubatedwithamixtureofanti-histone-biotinandanti-DNA-peroxidase.Duringtheincubationinterval,nucleosomeswillbecapturedviatheirhistonecomponentbytheanti-histone-biotinantibody,whilebindingtothestreptavidin-coatedmicroplate.Simultaneously,anti-DNA-peroxidasebindstotheDNApartofthenucleosomes.Afterremovaloftheunboundantibodies,theamountofperoxidaseretainedintheimmunocomplexisphotometricallydeterminedwithABTSasthesubstrate.

Figure1:SchematicshowingtheprincipleoftheCellDeathDetectionELISAPLUS.

TypicalExperiment

Figure2:Dose-dependentinductionofapoptosisinU937cells,detectedusingtheCellDeathDetectionELISAPLUS. U937cells(104cells/well,in200μl)wereincubatedwith differentconcentrationsofcamptothecin(CAM)for4hoursat37°C.Beforeandafterlysis,cellswerecentrifugedandthesupernatantwasanalyzed.Resultswereplottedasdosevs.response.
Result:Amountsofcytoplasmicoligonucleosomes(anindicatorofapoptosis)increaseasCAMconcentrationincreases.Cellculturesupernatantsremovedfromthecellsaftertreatment(butbeforelysis)gavenosignal,indicatingthattherewerenonecroticcellsduringthetreatment.

 

订购指南：以下产品部分现货供应

检测方法

产品名称

产品货号

规格

价格￥

FACS

Fluorescence

orlight

microscopies

Annexin-V-FLUOS

11828681001

250T

5023

Annexin-V-Alexa568

03703126001

250T

5443

Annexin-V-Biotin

11828690001

250T

5023

Annexin-V-FLUOSStainingKit

11858777001

50T

1932

Annexin-V-FLUOSStainingKit

11988549001

250T

6233

ELISA

Caspase3ActivityAssay

12012952001

96T

6578

CellDeathDetectionELISAPLUS

11774425001　

96T

3814

TUNEL

InSituCellDeathDetectionKit

Fluorescein

11684795910

50T

4662

InSituCellDeathDetectionKit

AP

11684817910

50T

5267

InSituCellDeathDetectionKit

POD

11684817910

50T

5267

InSituCellDeathDetectionKit

TMRred

12156792910

50T

3931

 

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