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Active Peptide-N4-N-Acetyl-Beta-D-Glucosaminyl Asparagine Amidase F (PNGaseF)
Ngl; PNG; PNGase F; Glycopeptide N-glycosidase; N-glycanase
- Product No.APX267Ge01
- Organism SpeciesPan-species (General) Same name, Different species.
- Buffer FormulationPBS, pH7.4, containing 0.01% SKL, 5% Trehalose.
- Traits Freeze-dried powder
- Purity> 90%
- Isoelectric Point8.0
- ApplicationsCell culture; Activity Assays.
- DownloadInstruction Manual
- UOM 10µg50µg 200µg 1mg 5mg
- FOB
US$ 288
US$ 720
US$ 1440
US$ 4320
US$ 10800
For more details, please contact local distributors!
ACTIVITY TEST
PNGase F (Peptide-N-glycosidase F) is a kind of enzymes for the deglycosylation of glycoproteins. The enzyme releases asparagine-linked oligosaccharides from glycoproteins and glycopeptides by hydrolyzing the amide of the asparagine (Asn) side chain. Thus, the activity of recombinant PNGase F measured by deglycosylating Interferon Gamma (IFNg) under denatured conditions. Prepare 10×denaturing buffer(5% SDS,400mM DTT), dilute IFNg to 1 μg/μl by 1×denaturing buffer,then heat the solution to 100℃for 10 minutes to denature the glycoprotein. Cool to room temperature and microcentrifuge briefly. Dobule dilution of recombinant PNGase F by assay buffer(50mmol/L Tris(pH7.4),1% NP-40), add 10 μl denatured IFNg to 10 μl different concentration of recombinant PNGase F, incubate reaction mixture at 37℃ for 1 hour. Stop the reaction by heating to 100℃ for 5 minutes, assess deglycosylation by SDS-PAGE. In the 10 μl reaction system, the amount of PNGase F needed to remove more than 95% carbohydrates from 10 μg denatured IFNg in 1 hour at 37℃ was defined as a unit. In this procedure, one unit is equal to 2.5ng recombinant PNGase F . The results are shown in Figure 1.
USAGE
Reconstitute in 10mM PBS (pH7.4) to a concentration of 0.1-1.0 mg/mL. Do not vortex.
STORAGE
Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.
STABILITY
The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.
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| Catalog No. | Related products for research use of Pan-species (General) Organism species | Applications (RESEARCH USE ONLY!) |
| RPX267Ge01 | Recombinant Peptide-N4-N-Acetyl-Beta-D-Glucosaminyl Asparagine Amidase F (PNGaseF) | Positive Control; Immunogen; SDS-PAGE; WB. |
| APX267Ge01 | Active Peptide-N4-N-Acetyl-Beta-D-Glucosaminyl Asparagine Amidase F (PNGaseF) | Cell culture; Activity Assays. |