View product documentation

Product guide

Certificate of stability

Product Specification:

LudgerZyme PNGase L is a recombinant glycoglycoamidase cloned from Flavobacterium akiainvivens.

Application:

LudgerZyme PNGase (LZ-PNGaseL-50-KIT) is suitable for the release of N-linked glycans in solution. The enzyme cleaves between the innermost GlcNAc of the oligosaccharide moiety at its attachment point to the asparagine residue on the protein and subsequently converts the asparagine into aspartic acid. Released glycans with free reducing termini can be labelled using LudgerTag labelling technology for fluorescence and high MS sensitivity detection.

Source: Flavobacterium akiainvivens.

Contents:

Each kit contains the following materials and reagents:

Suggested Usage:

Denaturing reaction conditions:

1. Make up the sample volume to 9 µL with ultrapure water.

2. Add 1 µL of 10X Denaturation Solution [LZ- 10X-DENAT-50] to each glycoprotein sample. Close the reaction vials, vortex thoroughly and briefly centrifuge to ensure the samples are completely dissolved.

3. Incubate the samples at 100 °C for 10 minutes.

4. Add 2 µL of 10X Reaction Buffer [LZ-10X-REACT-50] to each glycoprotein sample.

5. Add 2 µL of 10% NP-40 solution [LZ-NP40SOL-50].

6. Add 4 µL of water.

7. Add 2 µL of PNGase L [LZ-PNGaseL-50]. Close the reaction vials, mix gently and briefly centrifuge.

8. Incubate the samples at 37 °C for 1h.

No denaturing reaction conditions:

1. Make up the sample volume to 18 µL with ultrapure water.

2. Add 2 µL of 10X Reaction Buffer [LZ-10X-REACT-50] to each glycoprotein sample.

3. Add 2-5 µL of PNGase L [LZ-PNGaseL-50]. Close the reaction vials, mix gently and briefly centrifuge.

4. Incubate the samples at 37 °C for 4-24 h.

Specificity:

This enzyme is suitable for the release of a broad spectrum of N-glycans (high-mannose, hybrid and complex) with lower activity against bisecting GlcNAc-containing glycans compared to PNGaseF, from glycoproteins and glycopeptides, including those from non-mammalian sources such as plants, insects and parasites carrying α(1-3) linked core fucose and xylose moieties (see figures 1 and 2).

Number of Samples: The kit is sufficient for approximately 50 samples.

Amount of Sample: As a guideline, up to 100 μg of glycoprotein per sample.

Suitable Samples: Glycoproteins and glycopeptides containing N-linked glycans.

Storage: Store at 4°C. Protect from sources of heat and light.

Shipping: The product should be shipped at 4°C.

Handling: Ensure that any glass, plasticware or solvents used with this item are free of environmental carbohydrates and contaminating enzymes. Use powder-free gloves for all sample handling procedures and avoid contamination with environmental carbohydrates.

Safety: For research use only. Not for human or drug use

Please read the Safety Data Sheets (SDS's) for all chemicals used. All processes involving labelling reagents should be performed using appropriate personal safety protection – safety glasses, chemically resistant gloves (e.g. nitrile), lab coat, and, when appropriate, in a laboratory fume cupboard.