Gly-X™ N-Glycan Rapid Release and Labeling with APTS Express™ Kit (96-ct) [GX96-APTS]

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Description

The Gly-X™ N-Glycan Rapid Release and Labeling with APTS Express™ kit utilizes a novel in-solution enzymatic protein deglycosylation followed by Express labeling of released N-glycans with 8-aminopyrene-1,3,6-trisulfonic acid (APTS). With deglycosylation carried out in solution, the method is simple, rapid and suitable for automation. After a simple clean-up step, the APTS-labeled glycan samples are ready for analysis by CE, CE-MS, and other methods.

Sample prep time: ~2.5 hours. 

Other benefits include:

    • Reductive amination APTS Express labeling chemistry without dry down
    • 5-minute PNGase F digestion provides unbiased N-glycan release
    • Flexible, high-throughput format: process 1 to 96 samples
    • Optimized cleanup removes excess free dye, protein, and other interfering compounds.

    Contains:

      • GX96-100 Gly-X Deglycosylation Module (96-ct)
      • GX96-601 Gly-X APTS Express Labeling Module (96-ct)
      • GX96-602 Gly-X APTS Express Clean-up Module (96-ct)

      Equipment and reagents provided by user:

        • 96-well Thermocycler or two independent heat blocks, set for 90°C and 50°C
          • Thermocycler (Corning THERM-1001, 110V; THERM-1000, 230V)
          • Note: Two GlykoPrep Heaters, WS0271, can be fitted with VWR 13259-260 Modular Heating Blocks
        • Vacuum manifold (Millipore MSVMHTS00-HTS)
        • Vacuum pump (Millipore WP6211560, 110 V; WP6122050, 220V; Welch WOB-L Pump 2522)
        • ProZyme Vacuum Manifold Spacer (GX100, or GX400 Gly-X 2-AB/APTS Starter Kit - included free of charge with first Gly-X order, please contact us for details)
        • Acetonitrile (ACN), MS-grade (Fisher A955-4 recommended
        • DI Water

        Size: 1 kit (96-ct)

        Product Code: GX96-APTS

        Tech Info

        GX96-APTS Gly-X with APTS Express User Manual

        FAQs

        Q. Can I use more than the recommended upper limit of 40 µg protein per reaction?

        A. Maybe, although it will depend on the protein. You would have to test to ensure that labeled glycans from > 40 µg of your protein can be successfully loaded onto the Gly-X Cleanup Matrix without blockage. 

        Q. My samples are not loading completely onto the Gly-X Cleanup Matrix. What is happening?

        A. This may be caused by the nature of your protein sample, or by matrix effects caused by the composition of your formulation buffer. This can be addressed by using less protein per reaction, or by buffer exchanging your protein prior to starting the Gly-X protocol.

        Q: Can I use Eppendorf microtubes for the Denaturation and Digestion Steps, rather than a PCR plate?

        A: Yes, leave the tubes open for the heating steps, and ensure that material does not get in the lid during mixing.

        Q: Is it possible to use a plate shaker/heater such as the Eppendorf Thermo Mixer C formixing and heating during the Denaturation and Digestion protocols?

        A: Yes, this is possible. Mix rate should be high enough to vortex material in wells.

        Q: What is the Finishing Reagent for prior to the APTS labeling step?

        A: The Finishing Reagent is a weak acid, and drives conversion of the glycosylamine (-NH2) to a glycan with a free reducing end (-OH) prior to APTS labeling by reductive amination. 

        Safety

        Product Safety Documentation for GX96-APTS:

        Product/Part No. Description
        WS0343 Gly-X N-Glycanase (PNGase F)
        WS0344 Gly-X Digestion Buffer
        WS0358 Gly-X Denaturant
        WS0360 Gly-X APTS Reductant Solution
        WS0380 Gly-X APTS Solution
        WS0381 Gly-X APTS Catalyst
        WS0385 Gly-X APTS Finishing Reagent
        Certificates of Analysis

        This kit contains multiple modules, each with their own Certificate of Analysis, which may be searched for & accessed here.