商品编号


GKE-5006B



品牌


ProZyme



公司


ProZyme, Inc



公司分类


Glyko Enzymes



Size

200 mU

商品信息

Description

PNGase F, peptide-
N
-glycosidase F,
peptide-
N
4
-(
N-
acetyl-β-glucosaminyl)asparagine amidase.
?Releases intact N-glycans by cleaving
?between the innermost GlcNAc and Asn.?

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 and some insect 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-5006 N-Glycanase (PNGase F).
R
1
: N & C substitution other than H
R
2
:?may be H or the rest of an oligosaccharide
R
3
:?may be H or?
α(1-6)-linked fucose

Applications:


Release of intact N-linked glycans from glycopeptides and glycoproteins

Structure-function studies of N-glycosylated glycoproteins

Preparation of deglycosylated proteins for molecular weight estimation or crystallography studies


Supplied at a concentration of?≥ 2.5 U/ml. For a more concentrated formulation of?≥ 10?U/ml, which may be useful for native digestions, see
GKE-5010B
or
GKE-5020B
(EDTA-free).




"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.


Formulation:

20 mM Tris-HCl, 50 mM NaCl, 1 mM EDTA (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:
200 mU in 80 ?l

Concentration:
?≥ 2.5 U/ml

Product Code:
GKE-5006B

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.?
Int J Syst Evol Microbiol.
2005 May;55(Pt 3):1287-93.