LudgerZyme PNGase L Kit

Flavobacterium akiainvivens.

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

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.

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.