PNGase F-Fast 

PNGase F-Fast is a glycosidase that catalyzes the cleavage of internal glycosidic bonds in oligosaccharides. PNGase F-Fast removes almost all N-linked oligosaccharides from glycoproteins. PNGase F-Fast can release N-glycans from glycoproteins in the sugar analysis workflow. The cleavage site is: the glycosidic bond between the innermost N-acetylglucosamine and asparagine. PNGase F-Fast is an improved reagent that allows for rapid deglycosylation of antibodies and antibody fusions within minutes^[1][2].

One-step procedure
1. Dissolve 100 μg of antibody or glycoprotein in deionized water and dilute to 16 μL with deionized water;
2. Add 4 μL of reaction buffer (0.25 M sodium phosphate pH 7.50, 0.1 M DTT (HY-15917), 25 g/L Lauric acid (HY-Y0366) and gently pipette to mix.
3. Add 1 μL of PNGase F-Fast and gently pipette to mix.
4. Incubate at 50°C for 10 min.
5. After derivatization, N-glycans can be directly isolated and purified using solid-phase extraction for downstream glycoform analysis.

Two-Step Procedure
For some glycosylated antibodies, complete deglycosylation requires sample preheating. The steps are as follows:
1. Dissolve 100 μg of antibody or glycoprotein in deionized water and adjust the volume to 16 μL with deionized water;
2. Add 4 μL of reaction buffer (same composition as above) and gently pipette to mix;
3. Incubate at 80°C for 2 minutes, then cool on ice;
4. Add 1 μL of PNGase F-Fast and gently pipette to mix;
5. Incubate at 50°C for 10 minutes;
6. After derivatization, N-glycans can be directly isolated and purified using solid-phase extraction for downstream glycoform analysis.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

83534-39-8

Liquid

Colorless to light yellow

[PNGase F-Fast]

Room temperature in continental US; may vary elsewhere.

Please store the product under the recommended conditions in the Certificate of Analysis.

• [1]. Vilaj M, et al. Evaluation of different PNGase F enzymes in immunoglobulin G and total plasma N-glycans analysis. Glycobiology. 2021 Jan 9;31(1):2-7.  [Content Brief]

  [2]. Huang J, et al. Highly Efficient Release of Glycopeptides from Hydrazide Beads by Hydroxylamine Assisted PNGase F Deglycosylation for N-Glycoproteome Analysis. Anal Chem. 2015 Oct 20;87(20):10199-204.  [Content Brief]