N-Glycanase® (EDTA-Free), 1 U [GKE-5016D]

Price:

Description

PNGase F, peptide-N-glycosidase F, peptide-N4-(N-acetyl-β-glucosaminyl)asparagine amidase. Releases intact N-glycans by cleaving between the innermost GlcNAc and Asn. Same manufacturing process as N-Glycanase for GlykoPrep (EDTA-free), supplied at the GKE-5006 concentration of ≥ 2.5 U/ml.

Source:
Recombinant gene from Elizabethkingia meningosepticum, expressed in E. coli. The source organism was previously known as Chryseobacterium [Flavobacterium] meningosepticum [1]. 

Specificity: 
Cleaves all N-linked complex, hybrid or high mannose oligosaccharides, unless α(1-3) core fucosylated, as in plant glycans. Asparagine must be peptide-bonded at both termini. Phosphate, sulfate and sialic acid groups attached to the oligosaccharide do not affect cleavage. Endo F free.

 

Figure 1: Cleavage specificity for GKE-5016 N-Glycanase (PNGase F).
R1: N & C substitution other than H
R2: may be H or the rest of an oligosaccharide
R3: may be H or α(1-6)-linked fucose

"A & B" pack sizes ship with:

    • 1 ea WS0010 5x N-Glycanase Reaction Buffer
    • 1 ea WS0012 Denaturation Solution
    • 1 ea WS0013 Detergent Solution
    • 1 ea WS0145 5x N-Glycanase Tris Reaction Buffer (MS applications)

"D" pack size does not ship with the above buffers. Buffers are available with "D" pack size on request.

Formulation:
20 mM Tris-HCl, 50 mM NaCl (pH 7.5)

Reaction Buffer:
5X concentrated buffer, when diluted gives 20 mM sodium phosphate pH 7.5, containing 0.02% sodium azide.

MW:
~35,000 Daltons

pH optimum/range:
8.6 / 7.5 - 9.5

Unit Definition:
One unit is defined as the amount of enzyme required to catalyze the release of N-linked oligosaccharides from 1 μmole of denatured ribonuclease B per minute at 37°C, pH 7.5.

Not formulated for FACE® analysis, use GK50050 instead.

Size: 1 U in 400 µl

Concentration: ≥ 2.5 U/ml

Product Code: GKE-5016D

1. Kim KK, Kim MK, Lim JH, Park HY, Lee ST. Transfer of Chryseobacterium meningosepticum and Chryseobacterium miricola to Elizabethkingia gen. nov. as Elizabethkingia meningoseptica comb. nov. and Elizabethkingia miricola comb. nov.  2005 May;55(Pt 3):1287-93.

Tech Info

GKE-5016D Technical Data Sheet  

Safety

Product Safety Documentation for GKE-5016D:

Product/Part No. Description
GKE-5016D N-Glycanase® (EDTA - Free) 

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