商品介绍
| Description | |
|---|---|
| CatalogueNumber | APT500 |
| BrandFamily | Chemicon® |
| TradeName |
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| Description | CaspaTagPan-CaspaseInSituAssayKit,Sulforhodamine |
| Overview | Apoptosisisanevolutionarilyconservedformofcellsuicide,whichfollowsaspecializedcellularprocess.Thecentralcomponentofthisprocessisacascadeofproteolyticenzymescalledcaspases.Theseenzymesparticipateinaseriesofreactionsthataretriggeredinresponsetopro-apoptoticsignalsandresultinthecleavageofproteinsubstrates,causingthedisassemblyofthecell(Arends&Wyllie1991). CaspaseshavebeenidentifiedinorganismsrangingfromC.eleganstohumans.Themammaliancaspasesplaydistinctrolesinapoptosisandinflammation.Inapoptosis,caspasesareresponsIBLeforproteolyticcleavagesthatleadtocelldisassembly(effectorcaspases),andareinvolvedinupstreamregulatoryevents(initiatorcaspases).Anactivecaspaseconsistsoftwolargeandtwosmallsubunitsthatformtwoheterodimerswhichassociateinatetramer(Sleeetal.1999;Salvesen&Dixit1997;Alnemrietal.1996).Incommonwithotherproteases,caspasesaresynthesizedasprecursorsthatundergoproteolyticmaturation,eitherautocatalyticallyorinacascadebyenzymeswithsimilarspecificity(Sleathetal.1990). Caspaseenzymesspecificallyrecognizea4or5aminoacidsequenceonthetargetsubstratewhichnecessarilyincludesanasparticacidresidue.Thisresidueisthetargetforcleavage,whichoccursatthecarbonylendoftheasparticacidresidue(Thornberryetal.1992).Caspasescanbedetectedviaimmunoprecipitation,immunoblottingtechniquesusingcaspasespecificantibodies,orbyemployingFluorochromesubstrateswhichbecomefluorescentuponcleavagebythecaspase. TestPrinciple: CHEMICON"sInSituCaspaseDetectionKitsuseanovelapproachtodetectactivecaspases.ThemethodologyisbasedonFluorochromeInhibitorsofCaspases(FLICA).Theinhibitorsarecellpermeableandnon-cytotoxic.Onceinsidethecell,theinhibitorbindscovalentlytotheactivecaspase(Rotondaetal.1996).Thiskitusesasulforhodamine-labeledfluoromethylketonepeptideinhibitorofcaspase(SR-VAD-FMK),whichproducesaredfluorescence.Whenaddedtoapopulationofcells,theSR-VAD-FMKprobeenterseachcellandcovalentlybindstoareactivecysteineresiduethatresidesonthelargesubunitoftheactivecaspaseheterodimer,therebyinhibitingfurtherenzymaticactivity.Theboundlabeledreagentisretainedwithinthecell,whileanyunboundreagentwilldiffuseoutofthecellandiswashedaway.Thegreenfluorescentsignalisadirectmeasureoftheamountofactivecaspasepresentinthecellatthetimethereagentwasadded.Cellsthatcontaintheboundlabeledreagentcanbeanalyzedby96-wellplate-basedfluorometry,fluorescencemicroscopy,orflowcytometry. Application: TheCHEMICONInSituFLICAPan-CaspaseDetectionKitisafluorescent-basedassayfordetectionofactivecaspasesincellsundergoingapoptosis.Thekitisforresearchuseonly.Notforuseindiagnosticortherapeuticprocedures. |
| MaterialsRequiredbutNotDelivered | ·Culturedcellswithmedia ·Reagentstoinduceapoptosis ·15mLpolystyrenecentrifugetubes ·Ambervialsortubesforstorageof150Xconcentrateat-20°C ·600mLgraduatedcylinder ·Microscopeslides ·Hemocytometer ·Centrifuge(400xg) ·37°Cincubator ·Vortexer ·Adjustablevolumepipettorwithdisposabletips ·Deionizedwater ·PBS,pH7.4 ·DMSO |
| ProductInformation | |
|---|---|
| Components |
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| Detectionmethod | Fluorescent |
| StorageandShippingInformation | |
|---|---|
| StorageConditions | ·Storeunopenedkitmaterialsat2-8°Cuptotheirexpirationdate. ·ReconstitutedFLICAReagent(150X)shouldbefrozenat-20ºCforupto6months,andmaybethawedtwiceduringthistime.Aliquotintoseparateambertubesifdesired.Protectfromlightatalltimes. ·Storediluted(1X)washbufferupto-20ºCfor2weeks. |
| Applications | |
|---|---|
| Application | TheInSituFLICAPan-CaspaseDetectionKitforflowcytometryisafluorescent-basedassayfordetectionofactivecaspasesincellsundergoingapoptosis. |
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| 产品名 | 货号 | 公司分类 | 商城分类 | 美金价 |
| Millipore/475855-1R神奇滤布 密理博Millipore现货促销 | 475855-1R | Life Science Research | 膜 | 93.50 | Millipore/17-701/EZ-Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit | 17-701 | Life Science Research | 定量试剂盒 | 653.00 | Millipore/RIPAb+ Ago2 - RIP Validated Antibody and Primer Set | 03-110 | Life Science Research | 抗体套装 | 433.00 | Millipore/CBL468F | Anti-PECAM-1 Antibody, clone HC1/6, FITC conjugated/CBL468F/100 assays | CBL468F | Antibodies and Assays | 功能性抗体 | 298.00 | Millipore/ECM460 | αVβ3/PECAM Investigator Kit/ECM460/1 kit | ECM460 | Antibodies and Assays | 夹心法ELISA | 477.00 |
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