Cytotoxicity Detection KitPLUS (LDH)

CytotoxicityDetectionKitPLUS(LDH) 
Application
TheCytotoxicityDetectionKitPLUS(LDH)isafast,sensitive,andsimplemethodtoquantitatecytotoxicity/cytolysisbasedonthemeasurementofLDHactivityreleasedfromdamagedcellsusingthe96-wellor384-wellplateformat.Thekitcanbeusedinmanydifferentinvitrocellsystemswhendamagetotheplasmamembraneoccurs.Forexample:
Detectionandquantificationofcell-mediatedcytotoxicity. 
Determinationofmediator-inducedcytolysis. 
Determinationofthecytotoxicpotentialofcompoundsinenvironmentalandmedicalresearch,andinthefood,cosmetic,andpharmaceuticalindustries. 
Determinationofcelldeathinbioreactors.
Benefits
Suitableforhighthroughput:Fewerhandlingsteps.Notransfer,centrifugation,orprelabelingstepsarerequired. 
FlexIBLe:Definedassayconditionswhenusingastoppedcolorreaction. 
Safe:NorADIoactiveisotopesareused. 
Accurate:Resultsobtainedstronglycorrelatetolysedcellnumber.
Sensitive:Lowcellnumbersaredetected. 
Fast:TheuseofamultiwellELISAreaderpermitsalargernumberofsamplestobeprocessed.
ProductDescription
TheCytotoxicityDetectionKitPLUS(LDH)canbeperformedinahomogeneousformatandrequiresnotransferand centrifugationstepstoseparatethesupernatantfromthecells.Thecolorreactioncanbestoppedfordefinedassayconditions.
Samplematerial:Cellculturesgrownin96-or384-wellplatescanbemeasureddirectly.Aliquotsfromculturesgrowninotherformatscanbetransferredinto96-or384-wellplatesformeasurementwithoutremovingthecells.
Assaytime:10to30minutesforincubation.
Sensitivity:Lessthan100lysedcellscanbedetectedina96-wellplate.
BackgroundInformation
Celldeathisclassicallyevaluatedbythequantificationofplasma-membranedamage.Widelyusedmethodsarebasedonthedifferentialuptakeorexclusionofdyes,suchastrypanblueandpropidiumiodide,followedbycountingusingamicroscope.Themajordisadvantageisthatthesemethodsdonotpermit processingoflargesamplenumbers.Anothergroupofassaysisbasedonthereleaseofradioactiveisotopessuchas[51Cr]and[3H]-thymidineorfluorescencedyesfromprelabeledtargetcells.Thedisadvantagesoftheseassaysare(i)theuseofradioactiveisotopesinmostofthem,(ii)thenecessityforprelabelingofthetargetcells,and(iii)thehighspontaneousreleaseofmostlabelsfromtheprelabeledtargetcells.
Athirdtypeofassayisbasedonthemeasurementofcytoplasmicenzymeactivityreleasedbydamagedcells.Enzyme-releaseassayshavebeendescribed(e.g.,foralkalineandacidphosphatase);however,theiruseishamperedbythelowamountofrelevantenzymesthatispresentinmanycells,andbytheelaboratekineticassaysrequiredtoquantitatemostenzymeactivities.Incontrast,lactatedehydrogenase(LDH)isastablecytoplasmicenzymethatispresentinallcells.Itisrapidlyreleasedintothecell-culturesupernatantupondamageoftheplasmamembrane.
Contents
Catalyst(Diaphorase/NAD+mixture) 
DyeSolution(INTandsodiumlactate) 
LysisSolution 
StopSolution