Description
CatalogueNumber	APT116
BrandFamily	Chemicon®
TradeName	Apo-BrdU Chemicon
Description	Apo-BRDUIHCKit
Overview	TheCHEMICONAPO-BRDU-IHC™KitisatwocolorstainingmethodforlabelingDNAbreaksandtotalcellularDNAtodetectapoptoticcellsbyimmunohistochemistry(Lietal.1995).Thekitcontainstheinstructionsandreagentsrequiredformeasuringapoptosisincellsincluding:positiveandnegativecontrolslidesforassessingreagentperformance;reactionandblockingbuffersforprocessingindividualstepsintheassay;terminaldeoxynucleotidyltranferaseenzyme(TdT),bromodeoxyuridinetriphosphate(Br-dUTP),biotin-labeledanti-BrdUantibodyforlabelingDNAstrandbreaks,HRP-streptavidinconjugate,DAN,H2O2/Ureatabletsforcolorgeneration,andmethylgreensolutionforcounterstainingthecells.
MaterialsRequiredbutNotDelivered	1.Xylene-100% 2.Ethanol-100%,90%,80%,70%v/vwithdH2O 3.Methanol-100% 4.HydrogenPeroxide-30% 5.PBSorTris-bufferedsaline(1XTBS,20mMTrispH7.6,140mMNaCl) 6.10mMTris,pH8.0 7.1mMMgSO4in1XTBS(foruseingeneratingpositivecontrol) 8.DNaseI(foruseingeneratingpositivecontrol) 9.1-4%FormaldehydeinPBS,pH7.4 10.DistilledWater 11.Coplinjarswithslideholders 12.Beakerforrinsingslides 13.Slidehumidifyingchamber 14.Glasscoverslips 15.Mountingmedia 16.Microscope 17.Micro-pipettorwithdisposabletips 18.Microcentrifugetubes 19.Parafilm®(RegisteredtrademarkofAmericanNationalCanCompany) 20.Absorbentwipes 21.Crushedice

ProductInformation
Components	Positive/NegativeControlSlides,Naturalbox Note:2slides,eachcontainingamixturepositiveandnegativecontrolcells. BlockingBuffer,Whitecap H2O2/UreaTablets,Ambervial ProteinaseK,Pinkcap DABTablets,Ambervial TdTEnzyme,Yellowcap Br-Dutp,Violetcap 200XStreptavidin-HRPConjugate,Blackcap 5XReactionBuffer,Greencap Biotin-labeledBrdUmAb,Orangecap MethylGreensolution,Naturalcap, TheAPO-BRDU-IHC™Kitisshippedinonecontainerandshouldbestoredat-20°Cuponarrival. Note:Afterthekitisdefrostedbeforefirstuse,boththe5XReactionBufferandtheBiotin-labeledanti-BrdUantibodyshouldbestoredat2-8°C.Also,theMethylGreensolutionshouldbestoredatroomtemperature.DONOTRE-FREEZEanyofthesecomponentsoncethawed. Reagentbottleshavecolorcodedcapstoaidintheiridentification.Sufficientreagentsareprovidedtoprocess50slidesincluding2positive/negativecontrolslides.Thecontrolcellsarederivedfromahumanpromyelociticleukemiacellline(HL60)andincubatedwith0.5mg/mLactinomycinDfor19hourstoinduceapoptosis.
Detectionmethod	Chromogenic

StorageandShippingInformation

StorageConditions

PrecautionsandWarnings: 1.ThecomponentsofthiskitareforResearchUseonlyandarenotintendedfordiagnosticprocedures. 2.Afterinitialdefrosting,storethe5XReactionBufferandBiotin-labeledanti-BrdUantibodyat2-8ºC,andtheMethylGreensolutionatroomtemperature.DONOTRE-FREEZEanyofthesecomponentsoncethawed. 3.TdTEnzymewillnotfreezeat-20oC,becauseitisin50%(v/v)glycerolsolution. 4.The5XReactionbufferandBr-dUTPsolutioncontaincacodylicacidasabuffer.TheBiotin-labeledantibodycontains0.05%sodiumazideasapreservative.TheDABtabletscontaindiaminobenzidine.Thesematerialsareharmfulifswallowed.Avoidskincontactwithanyofthesecomponents;washimmediatelywithwater. 5.Uponwarmingthecomponents,centrifugethetubesfor30secondstoforcealltheliquidtothebottomofthetube. 6.Gloves,labcoatandprotectiveeyewearshouldbewornwhilestaining.

Applications
Application	TheAPO-BRDU-IHCKitisatwocolorstainingmethodforlabelingDNAbreaks&totalcellularDNAtodetectapoptoticcellsbyIHC.
KeyApplications	Immunohistochemistry

BIOLOGicalInformation
SpeciesReactivity	All

PhysicochemicalInformation

Dimensions

MaterialsInformation

MaterialsInformation

Millipore 提供三种不同的 Immobilon PVDF 膜，是针对不同的蛋白质印迹应用的理想选择。 现在我们还 提供采用按规格裁切的印迹方片膜和两层过滤纸组成的印迹三明治。   与硝酸纤维素膜相比，Immobilon 膜具有多种优点。 它们不易破裂或卷 曲，便于裁剪且在切割过程中不会碎裂。 它们还有低背景、广泛溶剂相容性以及优异的着色能力。 此外，它们可以重复检测。 Immobilon-P 膜（沃比森-供应） ·         0.45μm 孔径 ·         建议用于大多数免疫印迹，特别在蛋 白质大于 20kDa 时 ·         Millipore 的快速免疫检测实验方法不需要膜封闭并缩短洗涤时间，将检测时间至少减少了两个小时，而不致降低灵敏 度或特异性 用于高通量实验室的印迹三明治 ·         当您需要处理多个印迹时，印迹三明治是一种便利省时的选择。 我们的印迹三明 治包含多片 Immobilon-P 膜，其中插入了按规格裁切的多张层析级印迹纸。 转印滤纸 ·         按照大多数免疫印迹尺寸进行裁切的层析级转印滤纸。 Immobilon-PSQ 膜（沃比森-供应） ·         0.2μm 孔径和大的内表面面积 ·         较其它膜具有更高的 蛋白质吸附率和测序得率 ·         建议用于小于 20kDa 的印迹蛋白质 ·         防止小分子量的蛋白质的渗漏 Immobilon-FL 膜（沃比森-供应） ·        针对荧光应用进行优化的转印膜 ·         极低的背景提高了所 有荧光检测实验的灵敏度 ·         与所有常用的荧光探针（所有激发和发射波长）兼容 ·         是多态性检测和化学荧光应用的理想选择