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| Product Name | O-Glycosidase, Glycerol-Free, His-Tagged |
| Catalog No. | IVDRM-0305 |
| Description | O-Glycosidase, also known as endo-alpha-N-acetylgalactosaminidase, is a recombinant glycosidase expressed in E. coli with a gene originally derived from Enterococcus faecalis. Following removal of terminal sialic acids with a sialidase preparation, the enzyme catalyses cleavage of core 1 and core 3 O-linked disaccharides from glycoproteins. The storage buffer is deliberately formulated without glycerol, which favours optimal performance in HPLC workflows and mass spectrometric analysis. The enzyme carries a His tag, so after the digestion is complete it can be removed conveniently from the reaction by Ni2+ affinity chromatography. |
| Intended Use | Removal of core 1 and core 3 O-linked disaccharides from glycoproteins to enable downstream glycan and glycoprotein analysis. |
| Principle / Technology | After terminal sialic acids are trimmed with a sialidase, O-Glycosidase hydrolyses the glycosidic bond of core 1 and core 3 O-linked disaccharides, liberating intact O-linked disaccharides from the glycoprotein backbone. |
| Detection Method | O-linked core 1 and core 3 disaccharides on glycoproteins |
| Sample Type | Glycoproteins and glycopeptide preparations |
| Performance Range / Specifications | Supplied at 50 U per uL with purity above 90 percent by SDS-PAGE; the glycerol-free buffer system supports clean HPLC and mass spectrometry readouts. |
| Specificity | Acts on core 1 and core 3 O-linked disaccharide structures; sialic acids must be removed beforehand for efficient cleavage. |
| Reaction Conditions / Protocol | Denature the glycoprotein with the supplied 10x denaturation buffer, then incubate with O-Glycosidase in the 10x reaction buffer together with the provided detergent solution until O-linked glycans are released; deproteinise through a Ni2+ affinity step to clear the His-tagged enzyme. |
| Components / Formulation | O-Glycosidase at 50 U per uL; 10x glycoprotein denaturation buffer; 10x glycosidase reaction buffer; 10 percent detergent solution (NP-40 type). Volumes scale with the selected pack size. |
| Storage Conditions | -25 to -15°C |
| Shelf Life | 12 months |
| Package Specifications | 2500 U or 12500 U packs including matched buffer components |
| Product Form | Liquid enzyme in a glycerol-free buffer system, supplied with reaction buffers |
| Quality Control | Purity verified by SDS-PAGE (greater than 90 percent); activity confirmed with a sialylated FAM-labelled glycopeptide substrate before release. |
| Key Features | Glycerol-free formulation for HPLC and MS compatibility; His tag allows post-reaction enzyme removal by Ni2+ affinity; specific for core 1 and core 3 O-linked structures. |
| Purity | >90% (SDS-PAGE) |
| Concentration | 50 U/uL |
| Activity / Unit Definition | One unit is the enzyme amount that cleaves 1 pmol of a sialidase-treated FAM-labelled glycopeptide within 1 hour at 37°C |
| Source / Origin | Recombinant expression in E. coli; gene from Enterococcus faecalis |
| pH Range / Optimal pH | 7.5 (storage buffer) |
| Shipping Conditions | Cold pack |
| Expiration Date / Stability | Stable for the full shelf life when held at the recommended storage temperature; avoid repeated freeze-thaw cycles. |
| Regulatory / Compliance | For research and diagnostic reagent use; manufactured under controlled conditions with lot-specific release testing. |
| Compatibility | Compatible with HPLC and mass spectrometry workflows; requires prior desialylation of the substrate by a sialidase. |
| Recommended Buffer System | 20 mM Tris-HCl, 50 mM NaCl, 1 mM EDTA, pH 7.5, glycerol-free |
| Application Notes / Precautions | Complete desialylation is required before digestion; mix thoroughly after buffer addition and avoid contamination with exogenous glycosidases. |
| Batch-to-Batch Consistency | Each lot is released with activity and purity documentation; lot number is printed on the vial label. |
| Delivery Standard | Cold pack dispatch by default |
| Turnaround Time | Standard dispatch |
| Readouts / Downstream Assays | SDS-PAGE, HPLC, mass spectrometry of released O-linked glycans |
For in vitro diagnostic use.
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