Biotium/Caspase8 IETDR110 Fluorometric HTS Assay Kit/300122/100assays

Caspase-8IETD-R110FluorometricHTSAssayKitisspecificallydesignedforHTS-basedassays.Thekitprovidesahomogenousassaysystemforfastandhighlysensitivedetectionofcaspase-8activitybyfluorescenceinenzymaticreactionormammaliancells.

Caspase-8IETD-R110FluorometricHTSAssayKitisspecificallydesignedforHTS-basedassays.Thekitprovidesahomogenousassaysystemforfastandhighlysensitivedetectionofcaspase-8activitybyfluorescenceinenzymaticreactionormammaliancells.Thefluorogenicsubstrate(Ac-IETD)₂-R110containstwoIETDtetrapeptidesandiscompletelyhydrolyzedbytheenzymeintwosuccessivesteps.CleavageofthefirstIETDpeptideresultsinthemonopeptideAc-IETD-R110intermediate,whichhasabsorptionandemissionwavelengthssimilartothoseofR110(λ_Ex/λ_Em=496/520nm)buthasonlyabout10%ofthefluorescenceofthelatter.HydrolysisofthesecondIETDpeptidereleasesthedyeR110,leadingtoasubstantialfluorescenceincrease.

Note:Whilecaspase-8preferentiallycleavestheconsensussequenceIETDcomparedtoothersubstratesequences,othercaspasessuchascaspase-3alsocancleaveIETDefficiently.Overlappingcaspasesubstraterecognitionlimitstheusefulnessofcaspasesubstratepeptidesfordistinguishingbetweendifferentcaspaseactivitiesincelllysates.

TheassaykitincludesAc-IETD-CHO,whichisacaspase-8inhibitorandcanbeusedasanegativecontrol.Also,R110isprovidedinthekitforgeneratingastandardcurve,whichcanbeusedforquantifyingcaspase-8activity.

KitComponents:

• Celllysis/assaybuffer
• Enzymesubstrate(Ac-IETD)2-R110
• EnzymeinhibitorAc-IETD-CHO
• R110

Features:

• HTS-compatible:Single-stephomogenousassayspecificallydesignedforHTS-baseddetection.
• Fast:Fastenzymekinetics.
• Sensitive:Theenzymaticreactionformsintenselygreenfluorescentrhodamine110(R110)product.