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MOLECULAR BIOLOGY
PROTEIN BIOCHEMISTRY
GLYCOSIDASES
PNGase F (N-glycosidase F)

    PRODUCTS NAME                                                          CAT #              QUANTITY                      PRICE (W)    MORE INFORMATION  

PNGase F (N-glycosidase F)
EBG-1005
50,000 unit (500 unit/㎕)
300,000
DESCRIPTION
PNGase F (N-Glycosidase F) is a recombinant glycosidase expressed and purified from E.coli which carries the Flavobacterium meningosepticum gene.

PNGase F cleaves between the innermost N-acetylglucosamine and asparagine residues of the chitobiose core of N-linked glycans, leaving only asparagine.

PNGase F has a molecular weight of ~36.8 kDa.
The optimal pH is 7.5.
Substrate Specificity- Asn should be linked to at least one amino acid at the N- or C-terminal- hybrid N-linked oligosaccharides (except 1-3 linked fucose)

Product Components
- PNGase F : 100 ㎕ (500 unit/㎕)
- 10x Reaction Buffer : 500 mM sodium citrate, pH 7.5.

Unit Definition
- One unit is defined as the amount of enzyme required to remove > 95% of the carbohydrate from 10 ㎍ of denatured RNase B for 1 hr at 37°C in a total reaction volume of 10 ㎕.

Specific Activity
- 500 unit/㎕, 1,000 unit/㎍

Storage Buffer
- 20 mM Tris-HCl, pH 7.5, 50 mM NaCl, 5 mM EDTA, 50% glycerol.

PNGase F Reaction Condition
- 10 ㎍ of RNase B are denatured with 0.5% SDS, 1% β-mercaptoethanol at 100°C for 10 min. After the addition of Reaction Buffer to final 1x, PNGase F is added and the reaction mix is incubated for 1 hr at 37°C. Separation of reaction products is analyzed by SDS-PAGE (cleaved RNase B migrates faster). The activity of PNGase F is not affected by addition of 0.5% SDS. For a native glycoprotein, 5-fold more enzyme and longer incubation time will be required.

Storage Temperature
- Store at -20℃.

QC Tests
- Activity, SDS-PAGE purity, performance tests.

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