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| Product Name | Endoglycosidase H with Polyhistidine Tag, 1000000 U/mL, Glycerol-Free Buffer (10000 U / 50000 U) |
| Catalog No. | IVDRM-0301 |
| Description | Where high-mannose and some hybrid oligosaccharides have to be removed without disturbing complex N-glycans, endoglycosidase H is the usual answer. This enzyme is cloned from Streptomyces plicatus and expressed recombinantly in yeast with a polyhistidine tag for later removal, at a specific activity of 1000000 units per millilitre. It cuts the chitobiose core shared by high-mannose and hybrid chains, leaving the innermost N-acetylglucosamine on the protein. The point of difference here is the buffer: no glycerol is used, which avoids the interference glycerol causes in HPLC separations and in mass spectrometric detection, so this is the right choice when the digest goes directly into one of those methods. |
| Intended Use | Selective removal of high-mannose and some hybrid N-glycans before HPLC, mass spectrometry or general deglycosylation work on antibodies and other glycoproteins. |
| Principle / Technology | The endoglycosidase hydrolyses within the chitobiose core of high-mannose and hybrid structures, releasing the bulk of the glycan while one N-acetylglucosamine stays attached. |
| Sample Type | Glycoproteins, particularly those carrying high-mannose or hybrid N-glycans. |
| Performance Range / Specifications | 1000000 units per millilitre; glycerol-free buffer protects chromatographic and mass spectrometric performance; polyhistidine tag permits removal after digestion; unit definition anchored to denatured RNase B. |
| Sensitivity / LOD | Does not apply; sensitivity comes from the downstream detection chemistry. |
| Specificity | Theoretical molecular weight 29 kDa with an apparent 60 kDa band on unboiled SDS-PAGE; specific activity 1000000 units per millilitre; supplied as 10000 U or 50000 U with two 10x buffers. |
| Reaction Conditions / Protocol | Denature and treat the glycoprotein with the supplied buffers, add the enzyme and incubate at 37 °C, then analyse by the chosen chromatographic or mass spectrometric method. |
| Components / Formulation | Enzyme solution plus two 10x reaction buffers. |
| Storage Conditions | Store at -25 to -15 °C. |
| Shelf Life | 1 year from receipt when stored as recommended. |
| Package Specifications | Supplied as 10000 U or 50000 U with matched buffer volumes. |
| Product Form | Liquid enzyme in glycerol-free buffer. |
| Quality Control | Each lot is calibrated by unit activity against denatured RNase B and checked for absence of other glycosidase activity. |
| Key Features | - 1000000 units per millilitre for high capacity digestion - Glycerol-free buffer keeps HPLC and mass results clean - Selective for high-mannose and some hybrid N-glycans - Unit activity defined against denatured RNase B - Polyhistidine tag allows removal after the reaction |
| Purity | Recombinant glycosidase with no detectable contaminating protease. |
| Concentration | 1000000 units per millilitre. |
| Activity / Unit Definition | One unit removes more than 95% of carbohydrate from 10 ug of denatured RNase B within one hour at 37 °C in a 10 uL reaction. |
| Molecular Weight | Theoretical 29 kDa; about 60 kDa on unboiled SDS-PAGE and about 29 kDa after boiling. |
| Source / Origin | Cloned from Streptomyces plicatus and expressed recombinantly in yeast. |
| pH Range / Optimal pH | Storage and dilution buffer is 20 mM Tris with 50 mM sodium chloride and 5 mM EDTA at pH 7.5. |
| Shipping Conditions | Shipped cold. |
| Expiration Date / Stability | Stable across the stated shelf life when held at the recommended temperature and kept sealed. |
| Regulatory / Compliance | Supplied for laboratory research purposes. |
| Compatibility | Especially well suited to HPLC and to mass spectrometric analysis because no glycerol is present in the storage buffer. |
| Recommended Buffer System | 20 mM Tris, 50 mM sodium chloride, 5 mM EDTA at pH 7.5, free of glycerol. |
| Application Notes / Precautions | Do not boil samples if the unboiled form of the enzyme needs monitoring; note that the apparent molecular weight differs between boiled and unboiled preparations, which is normal. |
| Batch-to-Batch Consistency | Each production lot is released against fixed acceptance criteria so performance stays consistent between orders. |
| Model Type | Selective high-mannose glycan removal enzyme |
| Starting Material / Tissue Site | Recombinant glycosidase expressed in yeast with a polyhistidine tag |
| Delivery Standard | Enzyme vial plus two buffer vials |
| Turnaround Time | No lead time is published on the product page; availability is confirmed when the order is placed. |
| Readouts / Downstream Assays | Mass loss corresponding to removed high-mannose chains and the resulting glycan map. |
For in vitro diagnostic use.
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