Description:

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EndoD,alsoknownasEndoglycosidaseD,isarecombinantglycosidase,whichcleaveswithinthechitobiosecoreofpaucimannoseN-linkedglycans,withorwithoutextensionsintheantennae.EndoDistaggedwithachitinbindingdomain(CBD)foreasyremovalfromareaction,andissuppliedglycerol-freeforoptimalperformanceinHPLCandMS-intensivemethods.
ProductSource
AtruncatedEndoDgeneclonedfromStreptococcuspneumoniaeandexpressedinE.coliasafusiontochitinbindingdomain(2)Specificity
DetailedSpecificityFigure1:DetailedspecificityofEndoD
The?reactioncontained0.08mUofα1-2Mannosidase(ProZyme?#GKX-5009),50unitsofEndoDand2μl0.5?MNa0AcpH5.0bufferinatotalreactionvolumeof10μl.?Reactionswereincubatedat37°Cfor2hours.Following?removalofthebottombranch,EndoDisactive?onacomplexupperarm(1).ReagentsSupplied
Thefollowingreagentsaresuppliedwiththisproduct:
| Storeat(°C) | Concentration |
GlycoBuffer2 | -20 | 10X |
DTT | -20 | 10X |
Notes:
Todeglycosylateanativeglycoprotein,longerincubationtime,aswellasmoreenzyme,mayberequired.EndoDisnotrecommendedforusewithGlycoproteinDenaturingBuffercontainingbothSDSandDTT,asEndoDisinhibitedbySDSand,unlikeotherendoglycosidases,NP-40doesnotcounteracttheSDSinhibition.RemovalofEndoDfromthedeglycosylationreactioncanbescaleduplinearlywithlargeramountsofchitinmagneticbeads.ChitinMagneticBeadsBindingCapacityis0.4μg/μlofCBD-taggedprotein.50μlofchitinslurryaresufficienttoremove1-5μlofEndoD(50-250units)