Thefetuinglycoproteinstandardwasdevelopedforuseduringglycanreleaseandlabeling(particularlyforDMBsialicacidanalysis).Fetuinexistsinavarietyofglycoformscontainingbi-,tri-,andtetra-antennaryoligosaccharideswithvariablesialylation,aswellasO-linkedglycans.

AmountofGlycoproteinSupplied
GCP-FET-05500µg
GCP-FET-250U250µg
GCP-FET-50Ux450µgpervial–4vials

SourceFetalcalfserum

FormDry.Lyophilisedpowder.

MolecularWeight36kDa(proteinweightonly)

AmountofProtein
GCP-FET-05300µg
GCP-FET-250U150µg
GCP-FET-50Ux432µgpervial–4vials
IncomparisontoBSAstandard,determinedbyBCAassay.Valueroundedtonearestμg.

Analysis
FetuinglycanswerereleasedfromtheFetuinGlycoproteinStandard(Cat#GCP-FET-05)usingPNGase-F.

Followingreleasetheglycanswerelabeledusing2-Aminobenzamide(2-AB)usingtheLudgerTag2-ABGlycanLabelingKit(Cat#LT-KAB-A2).

Figure1:WatersBEHcolumnHPLCprofileof2-ABlabeledFetuinN-glycans,releasedbyPNGaseFfromGCP-FET-05batchB1B2-04.

Figure1showsaLudgerSepN2HPLCprofileofbovinefetuinN-glycans.TothoroughlyinvestigatetheN-glycanswefirstseparatethembasedonchargeonaLudgerSepC3column(Figure2)andthenruneachfractiononaLudgerSepN2column.

Figure2:LudgerSep-C3profileandsubsequentLudgerSepN2analysisofbovinefetuinPNGaseFreleasedN-glycansfromasimilarbatchofGCP-FET.
2-ABlabeledglycanswereseparatedontheLudgerSepC3columnandthesefractionswerethenseparatedontheLudgerSepN2column.ThisfiguredemonstratesthecomplexityofN-glycanspresentinthesample.AcombinationofLudgerSepC3/N2andexoglycosidasedigestionisrequiredtoidentifytheglycansandtheirrelativeabundance.N-neutralglycans,1S–monosialylatedglycans,2S–disialylatedglycans,3S–trisialylatedglycans&4S–tetrasialylatedglycans.Formoredetailedinformationonglycanidentification,pleaseseetheproductdocumentation.

StorageRefrigerate(-20°C)bothbeforeandafterdissolving.Thisproductisstableforatleast5yearsassupplied.

ShippingTheproductisshippedatambienttemperature.

HandlingOncedissolvedavoidrepeatedthawingandrefreezing,storageover3hoursatroomtemperatureorabove,exposuretolightandlongtermexposuretoacidasthesewillcauseglycandesialylation.

SafetyThisproductisnon-hazardousandhasbeenpurifiedfromnaturalsourcescertifiedtobefreeofallhazardousmaterialincludingpathogenicBIOLOGicalagents.