SmartEnzymes TransGLYCIT workflow

Hydrolysis of N-glycans

PNGase F (Peptide N-glycosidase F) is a glycoamidase hydrolyzing the amide bond between the polypeptide asparagine and the innermost GlcNAc of all mammalian asparagine-linked complex, hybrid, or high-mannose oligosaccharides.

The enzyme is widely used for sample preparation prior to MS analysis – to reduce the protein heterogeneity and enable released glycan analysis – and to study the functional role of the N-glycan.
 

  • Efficient hydrolysis of all mammalian N-glycans
  • Activity on a wide range of glycoproteins
  • Available immobilized in ready-to-use spin columns
  • Automation-friendly format for high-throughput N-glycan removal
N-linked glycans on glycoproteins
15+ min reaction time
No need for co-factors
Hydrolyzes the glycosidic bond between N-glycans and Asn

PNGase F Lyophilized

Lyophilized enzyme for hydrolysis of N-glycans on glycoproteins

PNGase F Immobilized

Immobilized enzyme for hydrolysis of N-glycans on glycoproteins in spin columns

PNGase F Automation

Lyophilized enzyme in 96-well plates for automated hydrolysis of N-glycans on glycoproteins

About PNGase F

PNGase F workflow

PNGase F is a glycoamidase hydrolyzing the amide bond between the polypeptide asparagine and the innermost GlcNAc of all mammalian asparagine-linked complex, hybrid, or high mannose oligosaccharides.

During the reaction, the asparagine residue from which the glycan is removed is deamidated to aspartic acid. The released oligosaccharide is left intact and can be used for further analysis. Removal of N-glycans is widely used for sample preparation for MS analysis to reduce the protein heterogeneity and enable released glycan analysis, and to study the functional role of the N-glycan.

The enzyme is expressed in E. coli from a recombinant gene derived from Elizabethkingia meningoseptica.

PNGase F Lyophilized

Lyophilized enzyme for hydrolysis of N-glycans on glycoproteins.

PNGase F Lyophilized is available as a lyophilized powder in 1000 or 5 x 1000 unit vials for digestion of 1 mg or 5 x 1 mg glycoprotein respectively.
 

  • Flexible format for method development
  • Possible to combine enzymes for improved efficiency
  • Amenable to automated workflows
  • Ready to use – just add water!

Available Products

PNGase F Lyophilized 1000 unitsLyophilized enzyme for hydrolysis of N-glycans on 1 mg glycoproteinG1-PF1-010520.00Buy / Request a Quote
PNGase F Lyophilized 5 x 1000 unitsLyophilized enzyme for hydrolysis of N-glycans on 5 x 1 mg glycoproteinG1-PF1-0502,080.00Buy / Request a Quote

For information on how to order, visit Place an Order or contact us directly at order@genovis.com.

Unit Definition

1 unit of PNGase F removes N-glycans from ≥ 95% of 1 µg etanercept when incubated in TBS buffer (50 mM Tris-HCl, 150 mM NaCl) pH 8.6 at 37°C for 1 h.

Content and Storage

PNGase F Lyophilized is supplied in 50 mM HEPES buffer pH 7.5, with no preservatives added. It is shipped cold and should be stored at -20°C upon arrival. After reconstitution, the enzyme is stable for at least 1 month at +4-8°C.

Instructions

Product Specifications

Safety Data Sheet

Certificate of Analysis

PNGase F Immobilized

Immobilized enzyme for hydrolysis of N-glycans on glycoproteins in spin columns.

The PNGase F Immobilized Microspin columns contain PNGase F enzyme covalently coupled to agarose beads, for hydrolysis of N-glycans on glycoproteins with reduced enzyme peaks in the final sample.

The glycoprotein sample is incubated with PNGase F Immobilized in the spin column, and the deglycosylated protein is then easily collected by a centrifugation step. The activity of PNGase F on some glycoproteins can be slow or inhibited due to steric hindrance - longer incubation times or denaturation of the glycoprotein may in these cases be required.

The Microspin columns are available for hydrolysis of 5 or 10 x 0.2 mg glycoprotein.
 

  • Easy-to-use spin columns
  • Increased enzyme-to-substrate ratio for improved digestion efficiency
  • Reduced enzyme peaks in the final sample

Available Products

PNGase F Immobilized Microspin 5 x 0.2 mgImmobilized enzyme for hydrolysis of N-glycans on 5 x 0.2 mg glycoprotein in spin columns under native conditionsG1-PF6-010578.00Buy / Request a Quote
PNGase F Immobilized Microspin 10 x 0.2 mgImmobilized enzyme for hydrolysis of N-glycans on 10 x 0.2 mg glycoprotein in spin columns under native conditionsG1-PF6-020988.00Buy / Request a Quote

For information on how to order, visit Place an Order or contact us directly at order@genovis.com.

PNGase F Immobilized Citations

Unit Definition

0.05 ml of settled agarose resin hydrolyzes ≥ 95% of N-glycans from 0.2 mg etanercept in TBS pH 8.6 in 1 h at 37°C as monitored by SDS-PAGE.

Immobilized PNGase F Microspin columns contain sufficient material to remove N-glycans from 0.2 mg glycoprotein.

Content and Storage

PNGase F Immobilized is supplied in 20% EtOH with no preservatives added, and is provided in packs of 5 or 10 columns. The columns are shipped cold, and should be stored at +4-8°C upon arrival. Do not freeze the product!

Instructions

Product Specifications

Safety Data Sheet

Certificate of Analysis

PNGase F Automation

Lyophilized enzyme in 96-well plates for automated hydrolysis of N-glycans on glycoproteins.

PNGase F Automation contains the PNGase F enzyme lyophilized in an automation-friendly 96-well plate format. It enables high-throughput N-glycan removal to facilitate simplified downstream analysis of a range of glycoprotein substrates. Depending on your workflow requirements and sample availability, PNGase F Automation is available lyophilized in 96-well plates for automated hydrolysis of N-glycans on 96 x 5 μg or 96 x 50 μg glycoprotein.
 

  • Enables automated high-throughput sample processing
  • Digestion of up to 96 samples in parallel
  • Two options available for digestion of different sample amounts
  • Ready to use – just add your sample!

Available Products

PNGase F Automation 96 x 5 unitsLyophilized enzyme in 96-well plates for automated hydrolysis of N-glycans on 96 x 5 𝜇g glycoproteinG1-PFA-005385.00Buy / Request a Quote
PNGase F Automation 96 x 50 unitsLyophilized enzyme in 96-well plates for automated hydrolysis of N-glycans on 96 x 50 𝜇g glycoproteinG1-PFA-050580.00Buy / Request a Quote

For information on how to order, visit Place an Order or contact us directly at order@genovis.com.

Unit Definition

One unit PNGase F removes N-glycans from ≥ 95% of 1 µg etanercept when incubated in TBS buffer (50 mM Tris-HCl, 150 mM NaCl) pH 8.6 at 37°C for 1 h.

Content and Storage

PNGase F Automation is supplied lyophilized in 50 mM HEPES buffer pH 7.5, with no preservatives added. The product is shipped at ambient temperature, and should be stored at -20°C upon arrival.

FAQ and Support

Popular FAQ

Yes! Contact us at info@genovis.com

No, it is not recommended. We can only guarantee optimal performance for one-time use.

The columns are recommended to be used for deglycosylation at native conditions only.

PNGase F Immobilized hydrolyzes the amide bond between the polypeptide asparagine and the innermost GlcNAc of all mammalian asparagine-linked complex, hybrid, or high mannose oligosaccharides. It does not remove N-glycans with α1-3 core fucosylation.

The recommended buffer for best performance of the column is TBS buffer at pH 8.6. Other buffers with neutral pH containing 0.15 M NaCl can also be used but optimization may then be required.

 

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