N-Glycan Kit (InstantPC-SPE)

Product Overview

Typical characterization of N-glycan profiles involves cleaving glycan chains with PNGase F, labeling with the derivatization reagentof2-AB, followed by analysis via high-performance liquid chromatography (HPLC). For the conventional 2-AB labeling workflow, enzymatic digestion duration and derivatization labeling time are the two primary rate-limiting steps. This rapid N-glycan analysis kit enables rapid enzymatic digestion, labeling and enrichment(higher fluorescence and mass spectrometry signals compared with 2-AB), with the entire sample pretreatment procedure taking less than 45minutes.

Key Features

Streamlined workflow

Integrates enzymatic digestion, labeling, and SPE-based enrichment/purification into a simplified three-step workflow.

Rapid enzymatic digestion

PNGase F digestion can be completed in approximately 5 minutes at 50 °C, helping reduce protein denaturation during processing.

Fast labeling reaction

InstantPC labeling is completed in approximately 5 minutes at room temperature under recommended conditions.

Efficient SPE-based enrichment

Solid-phase extraction (SPE) enables sample cleanup with a typical recovery rate of >90% and a processing time of ≤10 minutes per sample. When using a 96-well format combined with a vacuum manifold, throughput can be improved, with up to 12 samples processed in approximately 1.5 hours under optimized conditions.

Direct compatibility with LC/LC-MS analysis

Samples prepared using the SPE workflow can be directly used for downstream LC or LC-MS analysis without additional cleanup steps.

Applications

Rapid N-glycan profiling

Designed for fast preparation and analysis of N-linked glycans released from glycoproteins for LC or LC-MS-based characterization workflows.

High-throughput glycan sample preparation

Suitable for laboratories requiring time-efficient sample processing and streamlined N-glycan analysis workflows.

LC/LC-MS-based glycomics analysis

Applicable in liquid chromatography and mass spectrometry-based glycan profiling for research and analytical applications.

Performance Data

Performance Summary

No.	Performance	TestProtocol	Criteria		Results of Hzymes
No.	Performance	TestProtocol	Criteria		1	Blank Control	1. Prepare IgG samples and blank solution according to the IFU 2. No interfering peak shall appear at the retention time of the peak in IgG sample for the blank solution	No interfering peaks in blank solutions	YES
2	Repeatability	1. Prepare 6 parallel samples of IgG 2. Calculate the CV of G0F area & G0F% of the 6 samples	G0F	%Area CV ≤ 10%	1.36%
3	Intermediate Precision	1. Prepare 6 parallel samples of IgG by two persons separately 2. Calculate the CV of G0F area & G0F% of the 12 samples	G0F	%Area CV ≤ 10%	1.46%
4	Linearity	1. Prepare linear solutions of IgG between 1.0~10.0 mg/mL 2. Calculate the correlation coefficient R2 of sample concentration and G0F peak area	R2 of Area and concentration of G0F: R²≥0.990		0.9988
5	Inter-Batch Variation	1. Take 3 batches of reagents, and prepare 3 samples with 3 replicates per batch independently 2. Calculate the CV of G0F area & G0F% between batches respectively	G0F	Area CV ≤ 15%	2.48%
5	Inter-Batch Variation		G0F	%Area CV ≤ 10%	0.52%

Inter-Batch Variation

Linearity

FAQ

What should be adjusted if the protein amount needs to be changed?

Do not change the sample addition volume of 20μL, as this may affect the performance of the reaction system. Instead, 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 aliquoting is required, it should be divided immediately after reconstitution and 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 200μL. This can be adjusted according to specific sample conditions and instrument requirements. Optimization may be required depending on experimental setup.