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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

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!

Instructions

• PNGase F Lyophilized

Product Specifications

• PNGase F Lyophilized

Safety Data Sheet

• PNGase F Lyophilized

Certificate of Analysis

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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

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!

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!

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.

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