QA-Bio/Sialidase Sp/E-S007-200

Neuraminidase,N-acetylneuraminateglycohydrolase,Exo-alpha-sialidase

α(2-3)SialidaseSpcleavesexclusivelythenon-reducingterminalα(2-3)unbranchedsialicacidresiduesfromcomplexcarbohydratesandglycoproteins.Thereisnodetectableactivityonα(2-6)orα(2-8)linkagesoronbranchedα(2-3)linkages.Tocleaveallnon-reducingterminalsialicacidresiduesincludingbranchedsialicacids(linkedtoaninternalresidue)fromcomplexcarbohydratesandglycoproteins,useα(2-3,6,8,9)SialidaseAu(E-S001).

SourcerecombinantStreptococcuspneumoniaeinE.Coli

EC3.5.1.18

Contents
SialidaseSpin50mMsodiumphosphate,pH7.5

Includedwith20µLand60µLpacksizes:
5xReactionBuffer250mMsodiumphosphate,pH6.0

SpecificActivity≥250U/mg
Activity≥10U/ml

Molecularweight~75,000daltons

pHoptimum6.0

Suggestedusage1.Addupto100μgofglycoproteinor1nmolofoligosaccharidetotube.
2.Addwaterto14μL
3.Add4μL5XReactionBuffer.
4.Add2μLα(2-3)Sialidase.
5.Incubateat37°Cfor1hour

DesialylationmaybemonitoredbySDS-PAGEifthesizedifferentialbetweennativeanddesialylatedproteinissufficientfordetection.


SpecifictityCleavesthenon-reducingterminalalpha-(2-3)unbranchedsialicacidresiduesfromcomplexcarbohydratesandglycoproteins.

SpecificActivityDefinedastheamountofenzymerequiredtoproduce1µmoleofmethylumbelliferonein1minuteat37˚C,pH5.0fromMU-NANA[2′-(4-methylumbelliferyl)-alpha-D-N-acetylneuraminicacid].

StorageStoreenzymeat4˚C.

SialidaseSpReferences

Corfield,A.P.,H.Higa,J.C.PaulsonandR.Schauer.Thespecificityofviralandbacterialsialidasesforalpha(2-3)andalpha(2-6)-linkedsialicacidsinglycoproteins.BiochimBiophysActa744:121-126(1983).

Dwek,R.A.,C.J.Edge,D.J.Harvey,M.R.WormaldandR.B.Parekh.Analysisofglycoprotein-associatedoligosaccharides.AnnRevBiochem62:65-100(1993).

Kobata,A.Useofendo-andexoglycosidasesforstructuralstudiesofglycoconjugates.AnalBiochem100:1-14(1979).

Glasgow,LR.,J.C.PaulsonandR.L.Hill.SystematicpurificationoffiveglycosidasesfromStreptococcuspneumoniae.J.BiolChem252:8615-8623(1977).

Kelly,R.T.,D.GreiffandS.Farmer.NeuraminidaseactivityinDiplococcuspneumoniae.JBacteriol91:601-3(1965).

Prime,S.,J.Dearnley,A.M.Venton,R.B.ParekhandC.J.Edge.Oligosaccharidesequencingbasedonexo-andendoglycosidasedigestionandliquidchromatographicanalysisoftheproducts.JChromatogrA720:263-274(1996).