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Fast PNGase F Kit

Fast PNGase F Kit

Catalog Number: UA070119 Brand: UA BIOSCIENCE
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Regular price $544 USD
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Product Details

Product Specification


Stability & Storage

Store at -25 ~ -15℃ for 2 years

Reference


Background

PNGase F (N-Glycosidase F) is an asparagine amidase produced by Elizabethkingia miricola. PNGase F releases intact N-glycans by hydrolyzing the amide bonds of asparagine side chains (Asn-X-Ser/Thr, where X ≠ Pro), while converting Asn to aspartic acid (Asp). Unlike other glycosidases (such as Endo H), PNGase F can cleave all types of N-glycans (high-mannose, hybrid, and complex oligosaccharides), but it does not act on O-linked glycans or plant/insect N-glycans modified by core α-1,3-fucosylation. This enzyme is of significant importance in glycoprotein research, quality control in biopharmaceuticals, and proteomics. To enhance the convenience of experiments, we have developed the Fast PNGase F Kit, which simplifies the experimental procedures and reduces the reaction time to less than 20 minutes, thereby significantly improving experimental efficiency.

Components

Constituent

UA070119 -50 Rxns

UA070119 -100 Rxns

PNGase F

50 μl

 100 μl

5*Reaction Buffer

200 μl

400 μl


Protocol

1. Two -Step deglycosylation

1). Add the following components to one tube on ice:

Component

Volume

Final Concentration

Glycoprotein

-

10 μg/μl

5 × Reaction Buffer

2 μl

1 ×

ddH2O

Up to 10 μl

-

2). Denaturation: heating to 80℃ for 2 min

3). Deglycosylation: immediately after the reaction was completed, it was cooled on ice, and 1 μl of PNGase F was added and incubated at 50ºC for 10 minutes.

2. One -Step deglycosylation

1). Add the following components to one tube on ice:

Component

Volume

Final Concentration

Glycoprotein

-

10 μg/μl

5* Reaction Buffer

2 μl

1 ×

PNGase F

1 μl

-

ddH2O

Up to 10 μl

-

2). Deglycosylation: 1 μl of PNGase F was added and incubated at 50ºC for 10 minutes.

Guidelines

Two-step enzyme digestion is recommended to ensure higher enzyme digestion efficiency. 

This product is for scientific research purposes only. 

For your safety and health, please wear a lab coat and disposable gloves.

Picture

Bioactivity

Fast PNGase F Kit Deglycosylation Effect on RNase B. In a 10 μL reaction system, 10 μg of RNase B was added, incubated at 80°C for 2 minutes, cooled to room temperature, and then 1 U of PNGase F was added, followed by incubation at 50°C for 10 minutes. The following is the SDS-PAGE result:

M: Marker

Lane 1: Negative control (without PNGase F)

Lane 2: Competitor

Lane 3: UA070119

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