Recombinant F. meningosepticum PNGase F' (CystatinA Tag), CF

R&D Systems, part of Bio-Techne | Catalog # 7695-GHP

R&D Systems, part of Bio-Techne
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7695-GHP-020
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Key Product Details

Source

E. coli

Accession #

AAA85323 (PNGase F') & P01040 (Cystatin A)

Conjugate

Unconjugated

Applications

Enzyme Activity

View all PNGase F Proteins and Enzymes »

Product Specifications

Source

E. coli-derived f. meningosepticum PNGase F protein
Met 6-His tag Cystatin A
(Met1-Phe98)
Accession # P01040		 Linker PNGase F
(Ala41-Asn354)
Accession # AAA85323
N-terminus C-terminus

Purity

>90%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.

N-terminal Sequence Analysis

Met

Predicted Molecular Mass

48 kDa

SDS-PAGE

44-48 kDa, reducing conditions

Activity

Measured by its ability to deglycosylate ribonuclease B under denatured conditions.
>50% ribonuclease B (10 μg) is deglycosylated by 30 ng of Recombinant F. meningosepticum PNGase F' within 30 minutes, as measured under the described conditions.

Formulation, Preparation and Storage

7695-GHP
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and EDTA.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Background: PNGase F

PNGase F, peptide N-glycosidase F from Flavobacterium meningosepticum, catalyzes the hydrolysis of asparagine-linked high mannose, as well as hybrid and complex oligosaccharides from glycoproteins (1). Unlike glycosidases that hydrolyze glycosidic bonds, PNGase F is an amidase that cleaves the beta‑aspartylglucosamine bond between the innermost GlcNAc of N-glycans and asparagine residues of glycoproteins (2). The enzyme is highly active on various N‑glycans except those with the innermost GlcNAc modified with alpha1-3-linked core fucose, which is commonly found on plant Glycoproteins (3). Cleavage with PNGase F will convert the asparagine residue to an aspartic residue, allowing identification of the glycosylation sites by mass spectrometry (4). Recombinant PNGase F’ is a fusion of human cystatin A to PNGase F (Catalog # http://www.rndsystems.com/product_results.aspx?k=7695-GH">7695-GH) . PNGase F’ is a good alternative for PNGase F for deglycosylating proteins that have similar mass to PNGase F.

References

  1. Elder, J.H. and Alexander, S. (1982) Proc. Natl. Acad. Sci. USA 79:4540.
  2. Maley, F. et al. (1989) Anal. Biochem. 180:195.
  3. Tarentino, A.L. and Plummer, T.H. (1994) Methods Enzymol. 230:44.
  4. Zhang, H. et al. (2003) Nat. Biotechnol. 21:660.

Long Name

Peptide-N4-(N-acetyl-beta-D-glucosaminyl)asparagine Amidase F

Alternate Names

PNGF

UniProt

AAA85323 (PNGase F') & P01040 (Cystatin A)

Additional PNGase F Products

Product Documents for Recombinant F. meningosepticum PNGase F' (CystatinA Tag), CF

Product Datasheets

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Product Specific Notices for Recombinant F. meningosepticum PNGase F' (CystatinA Tag), CF

For research use only

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