CatalogueNumber | 2750 |
BrandFamily | Chemicon®
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TradeName | |
Description | BrdUCellProliferationKit |
Overview | TestPrinciple
WithCHEMICON?"sBrdUCellProliferationAssayKit,mousemonoclonalantibodyisusedtodetecttheBrdUinasample.Afteradditionofgoatanti-mouseIgG-peroxidaseconjugatedsecondaryantibody,substrateandstopsolution,theamountofBrdUisdetermined.ThehighertheOD,thehighertheBrdUconcentrationinthesample. |
MaterialsRequiredbutNotDelivered | 1.Pipettors&tipscapableofaccuratelymeasuring2-1000μL
2.Multi-channelorrepeatingPipettes
3.Washbottleormultichanneldispensorforwashing
4.2000mLgraduatedcylinder
5.PBS(137mMNaCl,2.7mMKCl,4.3mMNa2HPO4-7H2O,1.4mMKH2PO4)
6.DeionizedordistilledH2O.
7.96-wellmicroplatereaderwith450-540nmor450-595nmdualwavelengthor450nmsinglewavelengthfilter
8.Tissueculturemicroplate(96wellculturedish)
9.Sterilereagenttroughs
10.Microsyringefilter(0.2μm)
11.Syringe |
BackgroundInformation | Evaluationofcellcycleprogressionisessentialforinvestigationsinmanyscientificfields.Measurementof[3H]thymidineincorporationascellsenterSphasehasbeenatrADItionalmethodfordetectionofcellproliferation.Subsequentquantificationof[3H]thymidineisperformedbyscintillationcountingofautoradiography.Thistechnologyisslow,laborintensiveandhasseverallimitationsincludingthehandlinganddisposalofradioisotopesandthenecessityofexpensiveequipment.
Awellestablishedalternativeto[3H]thymidineuptakehasbeendemonstratedbynumerousresearchers.Inthesemethodsbromodeoxyuridine(BrdU),athymidineanalog,replaces[3H]thymidine.BrdUisincorporatedintonewlysynthesizedDNAstrandsofactivelyproliferatingcells.FollowingpartialdenaturationofdoublestrandedDNA,BrdUisdetectedimmunochemicallyallowingtheassessmentofthepopulationofcellswhicharesynthesizingDNA. |