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.

ProA is an upgraded version of 2-ABwithhigher fluorescence intensity and stronger mass spectrometry signals. This N-glycan analysis kit of ProA (magnetic bead method) enables rapid enzymatic digestion, labeling and enrichment, with the entire sample pretreatment procedure taking less than 60minutes.

Key Features

Streamlined workflow

Integrates enzymatic digestion, ProA labeling, and magnetic bead-based enrichment/purification into a simplified workflow covering three main steps.

Rapid enzymatic digestion

PNGase F digestion can be completed in approximately 10 minutes at 50 °C under recommended conditions.

Fast labeling reaction

ProA labeling can be completed in approximately 15 minutes at 65 °C. Compared with 2-AB,ProAprovides higher fluorescence intensity and stronger mass spectrometry signal response under comparable analytical conditions.

Magnetic bead-based enrichment and purification

Magnetic bead separation eliminates the need for lyophilization steps. Typical recovery rate is >90%, with a processing time of ≤15 minutes per sample.

Applications

Rapid N-glycan profiling

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

High-throughput glycan sample preparation

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

Fluorescence and MS-based glycan analysis

Applicable in glycan profiling workflows using fluorescence detection or mass spectrometry-based analytical platforms.

FAQ

How to deal with residual magnetic beads in samples after processing?

Magnetic beads may remain in the sample after purification. It is recommended to perform a centrifugation step using the matched filter tube prior to instrumental analysis (8,000 × g for 5 min, or adjust centrifugation time depending on the residual liquid volume).

For high-throughput workflows or larger batch processing, centrifugation time may be extended to more than 10 minutes to improve bead removal efficiency.

How should samples be handled before instrumental analysis to ensure stable results?

Since the final sample volume after dilution is relatively small and the buffer contains volatile acetonitrile, evaporation may affect peak area and peak shape. Therefore, it is recommended to minimize exposure time before analysis.

Best practice includes:

Injecting samples into the instrument immediately after centrifugation, or

Transferring samples to EP tubes and storing them in a sealed state (recommended storage time ≤ 3 days), or

Avoiding prolonged storage of samples in open vials prior to injection

Are there any recommendations for high-throughput analysis workflows?

For workflows involving a large number of samples, it is recommended to use a protective guard column (pre-column) to reduce contamination risk and extend the service life of the analytical column.