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Gibco™ PNGase F Glycan Cleavage Kit

Catalog No. A39245
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Catalog No. A39245 Supplier Gibco™ Supplier No. A39245
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Description

PNGase F Glycan Cleavage Kit includes all components necessary to perform the enzymatic removal of almost all N-linked oligosaccharides from glycoproteins.

Kit includes the PNGase F enzyme, which cleaves N-glycan chains at the innermost GlcNAc and asparagine residues of high mannose, hybrid, and complex oligosaccharides.

Features:

  • ≥95% purity
  • Recombinant PNGase F enzyme with no endoglycosidase F1, F2, or F3
  • Enzyme leaves N-glycan core oligosaccharides intact and suitable for downstream analysis
  • Suitable for use under native or denaturing conditions

Components included in the PNGase F Glycan Cleavage Kit are: 33,000 units of PNGase F (500,000 units/mL) and 1 mL PNGase F 10X buffer

Specifications

Content And Storage • 30,000 units PNGase F enzyme, recombinant (500,000 units/mL), store at -20°C
• 1 mL PNGase F 10X Buffer, store at -20°C
Shipping Condition Wet Ice
Form Liquid
Quantity 1 kit
Product Type PNGase F Glycan Cleavage Kit

Frequently Asked Questions (FAQs)

In the PNGase Glycan Cleavage Kit, can I use PNGase F under native or denaturing conditions?

Yes, PNGase F enzyme may be used in both native and denaturing conditions
In the PNGase Glycan Cleavage Kit, how much PNGase F should I use to remove my carbohydrate under native or DTT denaturing conditions?

When the protein is not denatured with SDS, PNGase F has to work harder to reach the cleavage site of the carbohydrate (due to secondary and tertiary protein structures). In these instances, using more enzyme and/or extended incubation times may help. These conditions will need to be determined empirically for your specific protein.
In the PNGase Glycan Cleavage Kit, is PNGase F compatible with downstream analysis such as HPLC and mass spectrometry?

PNGase F enzyme is stored in 50% glycerol. Therefore, HPLC and/or mass spectrometry is not compatible due to the fact that glycerol is not tolerated in instruments used for these applications.
In the PNGase Glycan Cleavage Kit, what are the typical reaction conditions for PNGase F, Recombinant?

Typical reaction conditions are as follows:

Optimal incubation times and enzyme concentrations must be determined empirically for a particular substrate.
1. Combine 1-20 µg of glycoprotein, 1 µL of 10X denaturing buffer (5% SDS, 400 mM DTT) and H2O (if necessary) to make a 10 µL total reaction volume.
2. Denature glycoprotein by heating reaction at 100 degrees C for 10 min.
3. Make a total reaction volume of 20 µL by adding 2 µL PNGaseF 10X Buffer, 1-5 µL PNGaseF enzyme, 2 µL 10% NP-40 and H2O.
4. Incubate reaction at 37 degrees C for 1 hr.
Note: Reactions may be scaled-up linearly to accommodate larger reaction volumes.
In the PNGase Glycan Cleavage Kit, what happens to the asparagine residue after PNGase F removes the sugar?

As PNGase F is a glycoamidase, the asparagine gets converted into aspartic acid.

For Research Use Only. Not for use in diagnostic procedures.