PNGase F
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PNGase F Immobilized
Immobilized enzyme for hydrolysis of N-glycans on glycoproteins in spin columns
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 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.
Why PNGase F Immobilized?
- 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 mg
Immobilized enzyme for hydrolysis of N-glycans on 5 x 0.2 mg glycoprotein in spin columns under native conditions
PNGase F Immobilized Microspin 10 x 0.2 mg
Immobilized enzyme for hydrolysis of N-glycans on 10 x 0.2 mg glycoprotein in spin columns under native conditions
For information on how to order, visit Place an Order or contact us directly at order@genovis.com.
Product Information
Activity
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!
Resources
Instruction for Use
Product Specifications
Safety Data Sheet
Certificate of Analysis
Find your certificate of analysis »Applications
Complete Fc N-glycan removal under native conditions, enabling simplfied subunit LC-MS analysis of therapeutic antibodies.
FAQ and Support
Does PNGase F Immobilized remove all N-linked glycans?
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.
Can the deglycosylation on the column be performed under denaturing conditions?
The columns are recommended to be used for deglycosylation at native conditions only.
Is it possible to purchase PNGaseF Immobilized in bulk?
Yes! Contact us at info@genovis.com
Which buffers can be used with the PNGaseF Immobilized column?
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.
Can I use FabRICATOR and PNGaseF in the same reaction?
Yes, PNGaseF and FabRICATOR can be used in a one-pot reaction as long as the reaction conditions follow the specifications for the FabRICATOR enzyme. It is important to have native reaction conditions for FabRICATOR to remain active.
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Genovis products are intended for research use only. They are not intended to be used for therapeutic or diagnostic purposes in humans or animals. All goods and services are sold subject to Genovis’ General Terms and Conditions of Sale.
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