Description
Thekithasbeendevelopedtoovercomethecommondifficultiesuniquetoneuronalsamplesincludingthefragilenatureofbraintissuesections,highbackgroundproblems,poorcounterstainingwithcommondyes,andtheneedtoperformduallabelingexperimentstodetectcellspecificantigensinconjunctionwithapoptoticcells.AkeyfeatureisNeuroPore™,aproprietarypermeABIlizationreagentthatgentlypermeabilizessampleswhileretainingcellmorphology.NeuroPore™alsocontainsblockingreagentstoallowitsuseasanantibodydiluentinimmunohistochemistryandtoreducebackgroundstaining.
DNAfragmentsgeneratedbyapoptosisareend-labeledwithmodifiednucleotidesusingahighlypurifiedterminaldeoxynucleotidyltransferaseenzyme(TdT).TheincorporatednucleotidesaredetectedusingahorserADIshperoxidasesystemthatcatalyzestheconversionofdiaminobenzidine(DAB)intoavisIBLedarkbrownprecipitate.NeuroTACS™IIcontainstheBlueCounterstaintoallowvisualizationofallcellswithinthesamplewithgoodcontrasttothebrownDABprecipitate.ADABenhancerisalsoprovidedwiththekitfortheoptiontointensifyordarkenDABstaining.Inaddition,theBlueCounterstainiscompatiblewiththered-coloredsubstratesusedforphosphatasedetectionindoublelabelingexperiments.Theprotocolincludesdetailsforlabelinginsituforapoptosisandantigendetectiononthesamesample.Toensurethatyourownsampleshavebeenprocessedandpermeabilizedcorrectly,weprovideTACS-Nuclease™,auniquereagentusedtogenerateapositivecontrolwithyourownsamples.Thisprovidesahighdegreeofconfidencefordatainterpretationandcanhelppinpointproblemstepsinthelabelingprocedure.
FEATURES:
- Uniquebuffersystemproducesmoreconsistentlabeling.
- Performancetestedonbrainsections.
- IncludesexclusiveNeuroPore™permeabilizationreagent.
- IncludesTACS-Nuclease™solutionforpreparingsample-dependentpositivecontrols.
- Helpsresolveuniqueproblemsencounteredwhendetectingapoptoticneuronalcells.
APPLICATIONS:
- Insitudetectionofapoptosisinfixedfrozen,paraffinembedded,orplasticembeddedcellsandtissues.
- Assistsintheidentificationofapoptoticmorphologies.
| Catalog#4823-30-Kincludes: | ||
| CatalogNumber | Description | Qty |
| 4800-30-01 | ProteinaseK | 1 |
| 4800-30-06 | Streptavidin-HRP | 1 |
| 4800-30-07 | Diaminobenzidine | 1 |
| 4800-30-09 | DABEnhancer | 1 |
| 4800-30-15 | TACS-Nuclease™ | 1 |
| 4800-30-16 | TACS-Nuclease™Buffer | 1 |
| 4810-30-02 | TACS®2TdTLabelingBuffer | 1 |
| 4810-30-03 | TACS®2TdTStopBuffer | 1 |
| 4810-30-04 | TACS®2TdTdNTP | 1 |
| 4810-30-05 | TdTEnzyme | 1 |
| 4810-30-14 | 50xManganeseCation | 1 |
| 4820-30-01 | NeuroPore™ | 1 |
| 4820-30-13 | BlueCounterstain | 1 |
DifferencesinthePhagocyticResponseofMicrogliaandPeripheralMacrophagesafterSpinalCordInjuryandItsEffectsonCellDeath,
AndrewD.GreenhalghandSamuelDavid
J.Neurosci.,Apr2014;34:6316–6322.
http://www.jneurosci.org/cgi/content/abstract/34/18/6316
HypoxiaInducibleFactor-1{alpha}(HIF-1{alpha})IsRequiredforNeuralStemCellMaintenanceandVascularStabilityintheAdultMouseSVZ
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SynapticSynthesis,Dephosphorylation,andDegradation:ANOVELPARADIGMFORANACTIVITY-DEPENDENTNEURONALCONTROLOFCDKL5
PaoloLaMontanara,LauraRusconi,AlbinaLocarno,LiaForti,IsabellaBarbiero,MarcoTramarin,ChetanChandola,CharlotteKilstrup-Nielsen,andNicolettaLandsberger
J.Biol.Chem.,Feb2015;290:4512–4527.
http://www.jbc.org/cgi/content/abstract/290/7/4512
