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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 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 terminus 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 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 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 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.

Number of Samples:

Vial contains 100 µl of solution. Enough for 50 samples if 2 µl is used for each reaction. 

Amount of Samples:

As a guideline up to 100 µg of glycoprotein per sample.

Suitable Samples:

Glycoproteins and glycopeptides containing N-linked glycans, including those from non-mammalian sources such as plants, insects, and parasites.

Storage:

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