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Native PNGase F (Peptide N Glycosidase F) [For Glycan Release] (CAT#: CB-P249-K) Datasheet

Product Type
Kit
Description
PNGase F is suitable for releasing all types (high mannose, hybrid and complex) N-linked glycans from glycoproteins and glycopeptides. PNGaseF does not remove the α(1-3) linkages commonly found on plant glycoproteins The core fucose is an oligosaccharide.
PNGase F cleaves N-linked (asparagine-linked) oligosaccharides from glycoproteins. The enzyme deaminates asparagine to aspartic acid, leaving oligosaccharides intact. Denaturation increases the cutting rate. Most natural proteins can still be completely N-deglycosylated, but the incubation time must be increased. The enzyme will remain fully active for at least 96 hours under the reaction conditions (37°C). PNGase F will not remove the oligosaccharides containing Alpha-(1,3)-linked core fucose that are commonly found on plant glycoproteins; for this, please use peptide N-glycosidase A.

There are many alternative enzymes that can be used to remove N-glycans, especially Endo F enzyme and EndoH. These enzymes cleave between the two N-acetylglucosamine residues in the oligosaccharide core, resulting in a truncated sugar molecule with an N-acetylglucosamine residue remaining on asparagine. This leaves the charged sugars, which help to retain the protein in the solution, which will precipitate after deglycosylation with PNGase F, thereby completely removing the oligosaccharides. Endo F1 cracks high mannose and some hybrid N-glycans. Endo F2 will eliminate dual antennas and high mannose (reduce the rate by 40 times). Endo F3 release of endotriol and fucosylated biantennary N-glycans. Endo H removes heterozygous or high mannose glycans.
Features
Contents: PNGase F in 20 mM Tris-HCl, pH 7.5 - 200µL
Specific Activity:>25 U/mg
Activity: 5 U/mL
Molecular weight: 36,000 daltons
pH range: 6-10, optimum 7.5
Source: Elizabethkingia miricola (was Chryseobacterium meningosepticum)
Size
0.3 U / 60 µL (Sufficient for up to 200 reactions)
Sample
Glycoproteins and glycopeptides containing N-linked glycans
Species Reactivity
Cleaves all asparagine-linked complex, hybrid or high mannose oligosaccharides unless alpha(1-3) core fucosylated; asparagine must be peptide bonded at both termini, Endo F free
Components
PNGase F in 20 mM Tris-HCl, pH 7.5 - 60µL
5x Reaction Buffer 7.5 - 250 mM sodium phosphate, pH 7.5
Denaturation Solution - 2% SDS, 1 M Beta-mercaptoethanol
Triton X-100 - 15% solution
Handling Advice
1. Add up to 200 µg of glycoprotein to an Eppendorf tube. Adjust to 35 µL final volume with de-ionized water.
2. Add 10 µL 5x Reaction Buffer 7.5 and 2.5 µL of Denaturation Solution. Heat at 100°C for 5 minutes.
3. Cool. Add 2.5 µL of Triton X-100 and mix.
4. Add 2.0 µL of enzyme to the reaction. Incubate 3 hours at 37°C.
Storage
Store enzyme at 4°C.
Shelf Life
12 months
Research Use
For research use only. Not for human or drug use

All products and services are for Research Use Only. Do Not use in humans.

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