The Krishzyme KriGly N-Glycan Kit utilizes a proprietary advanced rapid-tagging KriGly flourodye that delivers extreme mass spectrometry sensitivity enhancements. Our KriGly Fluorescent Dye defers from Rapifluor™ and ensures no active patent infringement while achieving identical performance. Rapid-tagging chemical analogs are engineered with strategic structural modifications in the non-reactive side chains – specifically altering the tertiary amine tail and linkers – while strictly preserving the core quinoline fluorophore.
MS Charge Enhancement: Ultra-Sensitive MS Detection: Delivers the exact same high-order mass spec sensitivity enhancement, allowing trace glycan characterization.
HILIC Retention Profile: Resolution Integrity: Maintains exceptional glycan peak separation across critical species (G0F, G1F, G2F) with minor gradient optimization.
The kit includes rapid enzymatic digestion using PNGase F, labeling with KriGly fluorophore, purification, and analysis by HPLC/LC-MS.
N-glycosylation significantly affects the biological function, half-life, immunogenicity, solubility, thermal stability, protease resistance, and aggregation of proteins. It is a key indicator of batch-to-batch consistency in antibody production. ICH-Q6B, various pharmacopoeias, and regulatory authorities have established quantitative requirements for N-glycan analysis.
Key Features:
Single Column / 96-well plate Workflows
Designed for 1 column / 96-well format processing, enabling parallel sample preparation and improved workflow efficiency in batch analysis.
Optimized 3-step Process
Integrates enzymatic digestion, labeling, and SPE-based enrichment/purification into a simplified workflow suitable for high-throughput applications.
Rapid Enzymatic Digestion
PNGase F digestion can be completed in approximately 5 minutes at 50°C, under recommended conditions.
Quick and High Sensitivity Labeling Reaction
Labeling reaction can be completed in approximately 5 minutes at room temperature. The Label Dye optimized for high sensitive applications equivalent to market competitors.
Efficient SPE-based Purification
Solid-phase extraction provides a typical recovery rate of >90%, with a processing time of ≤10 minutes per sample. When combined with a 96-well plate and vacuum manifold system, up to 12 samples can be processed in approximately 1.5 hours under optimized conditions.
Direct Compatibility with LC/LC-MS analysis
SPE-prepared samples can be directly used for LC or LC-MS analysis without additional purification steps.
Applications:
High-throughput N-Glycan Profiling
Designed for parallel preparation and analysis of N-linked glycans from glycoproteins in single column / 96-well format workflows.
Batch Sample Preparation for Glycan Analysis
Suitable for laboratories requiring efficient processing of multiple samples for LC or LC-MS-based glycomics studies.
LC/LC-MS-based Glycan Characterization
Applicable in liquid chromatography and mass spectrometry-based N-glycan analysis for research applications.
FAQs:
Do I adjust the protein concentration in the sample, while keeping the sample volume the same?
Yes, keep the sample addition volume of 20ul, as this may affect the performance of the reaction system. Should you desire, adjust the protein concentration to achieve the desired input amount.
How should the labeling reagent be handled after reconstitution?
The labeling reagent is recommended for immediate use after dissolution. If aliquots are required, do it immediately after reconstitution. The aliquots maybe stored at -80 °C for up to 5-7 days.
Is precipitation normal after adding acetonitrile?
Yes. The presence of slight precipitation after adding acetonitrile is normal and is caused by phase incompatibility between aqueous and organic solvents. This does not typically affect the reaction performance under recommended conditions.
Can the recommended sample volume be adjusted?
The recommended sample volume is 200ul. This can be adjusted according to specific sample conditions and instrument requirements. Optimization may be required depending on experimental setup.
