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PNGase F – CarboClip

Price range: 178,00€ through 800,00€

CarboClip is a recombinant peptide-N-Glycosidase F (PNGase F), from Elizabethkingia sp, overexpressed in E. coli that contains a hexahistidine tag in its C-terminal and cleaves between the GlcNAc and asparagine residues of high mannose, hybrid, and complex oligosaccharides from N-linked glycoproteins.

  • CarboClip is provided glycerol-free as a lyophilized powder containing Tris-HCl, NaCl, and Na2EDTA.
  • Unit Definition: One unit is defined as the amount of enzyme required to remove > 95% of the carbohydrate from 10 µg of denatured RNase B in 1 hour at 37°C in a total reaction volume of 10 µL.  
  • Unit Definition Assay: 10 µg of RNase B are denatured with glycoprotein denaturing buffer (0.5% SDS, 40 mM DTT) at 100°C for 10 minutes. After the addition of NP-40 and buffer (50 mM Sodium Phosphate pH 7.5), two-fold dilutions of PNGase F (1 μL) are added, and the reaction mixture is incubated for 1 hour at 37°C. The separation of reaction products is visualized by SDS-PAGE 12 % acrylamide and stained with Coomassie blue. 
  • Storage: Lyophilized powder: -80 °C and -20 °C >1 year. At room temperature for 10 days. Reconstituted protein should be stored at 4 °C and is stable for 2-3 months.
  • Glycosylation of Membrane Proteins: Removal of N-linked glycans from membrane protein samples using this enzyme has been described by Contessotto et al. Matrix Biol. 2019

Applications

  • N-glycomic analysis of therapeutic antibodies and therapeutic glycoproteins, as described by Sciex. 2019
  • Deglycosylation for Capillary Electrophoresis Separations of Glycans, as described by Mészáros et al.  Journal of Chromatography B. 2019.
  • Deglycosylation of Membrane proteins, as described by Contessotto et al. Matrix Biol. 2019.
  • Deglycosylation of Hemocyanins, as described by Jimenez et al. Front. Immunol. 2019.
  • Removal of N-Linked Glycans for Native Mass Spectrometry, as described by Bern et al. J. Proteome Res. 2018.
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Citations:

This PNGase F has been used in the following Scientific Publications:

Size

500U, 15000U, 75000U
PNGase F - CarboClip
PNGase F - CarboClip - Image 2