PNGase F

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  • Product overview
  • Product performance
  • Standard Operating Procedure
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  • Instructions
Product Nameicon
PNGase F
Catalog No.icon
Expression hosticon
Restriction sitesicon
PNGase F works by cleaving between the innermost GlcNAc and asparagine residues of high mannose, hybrid, and complex oligosaccharides from N-linked glycoproteins and glycopeptides. This results in a deaminated protein or peptide and a free glycan.
>95% as determined by SDS-PAGE quantitative densitometry by Coomassie Blue Staining.
1800000 U/mg
Unit Definitionicon
One unit of PNGase F will catalyze more than 95% of the deglycosylation of 10μg of denatured Ribonuclease B(RNase B) in 10 ul reaction system in one hour at 37°C.
Digestion reaction conditionicon
At 37 ℃, the enzyme digestion reaction 1h, every 1ug enzyme can digest 10ug substrate. (The mass ratio of enzyme to substrate is determined by our company based on the degree of glycosylation of the substrate used, for reference only)
20 mM Tris-HCl pH 7.5, 50 mM NaCl, 5 mM EDTA,50%Glycerol
Store at -20 °C for 12 months from the date of receipt.

1: Protein Deglycosylation Using Denaturing Conditions for SDS-PAGE a.Add up to 50μg of the target glycoprotein in water (or a compatible buffer at a low ionic strength) to a final volume of 12μl.

b. Add 1μl of 5% SDS.

c. Add 1μl of 1M DTT.

d. Denature sample by heating at 95°C for 5 minutes.

e. Cool sample for 5 minutes at room temperature.

f. Add 2μl of 0.5M sodium phosphate buffer (pH 7.5). Note: Other buffers can be used if they are within the acceptable pH range for PNGase F (pH 6–10).

g. Add 2μl of 10% NP-40. Note: Triton X-100 may be substituted for NP-40. H.Add 2μl of recombinant PNGase F. I.Incubate at 37°C for 1–3 hours.

2: Protein Deglycosylation Using Non-Denaturing Conditions for Mass Spectrometry

a. Add up to 20μg of glycoprotein in 50mM ammonium bicarbonate (pH 7.8) to a final volume of 18μl.

b. Add 2μl of recombinant PNGase F.

c. Incubate at 37°C for 2–18 hours.

Guarantee and Disclaimer:

After receiving the product, if finds that the product is mismatched, damaged or missing components, please keep the original package and submit the objection to the company by mail within seven working days. Failure to file an objection within the time limit is considered qualified.

When the buyer keeps the product, it should be kept in accordance with the storage conditions shown in the product label and the manual. If the product quality is caused by improper storage, it will not be guaranteed.

When the buyer tests the product, they should be submitted by the beginning of use when found the quality issue, rather than when the product was used or used up, in order to prepare our products for recycling and confirm the quality of the products. If it is the quality problem, our company is responsible for exchange or return. In the event of a claim, our company will compensate the discretion within the scope of the product price limit and will not accept any part of the value of the product itself. Since the date of receipt, the product has not been reflected for more than three months and should not be returned.

The products provided by our company are for research use only and should not be used for clinical diagnosis or treatment. If a unit or individual changes the use of our products without authorization, we will not bear any responsibility.