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362300 N-Glycosidase F, Chryseobacterium meningosepticum, Recombinant, E. coli

362300
Purchase on Sigma-Aldrich

Przegląd

Replacement Information

Products

Numer katalogowyOpakowanie Ilość/opak.
362300-1SET Butelka szklana 1 set
Description
OverviewRecombinant, Chryseobacterium meningosepticum N-Glycosidase F expressed in E. coli. This enzyme from Chryseobacterium (formerly Flavobacterium) meningosepticum catalyzes the hydrolysis of N-glycans from high mannose, hybrid and complex glycopeptides, and from glycoproteins generating asparagine-linked oligosaccharides.
Note: 1 mU = 1 milliunit.
Catalogue Number362300
Brand Family Calbiochem®
SynonymsPeptide-N-glycosidase F, Peptide-N⁴-(acetyl-β-glucosaminyl)-asparagine Amidase, PNGase F, Glycopeptidase F
References
ReferencesFan, J.-Q., and Lee, Y.C. 1997. J. Biol. Chem. 272, 27058.
Altmann, F., et al. 1995. Glycoconj. J. 12, 84.
Product Information
CAS number83534-39-8
Unit of DefinitionOne unit is defined as the amount of enzyme that will completely catalyze the release of N-linked oligosaccharides from 1 µmol denatured ribonuclease in 1 min at 37°C, pH 7.5.
EC number3.5.1.52
FormLiquid
FormulationEnzyme supplied in 50 mM NaCl, 20 mM Tris-HCl, 1 mM EDTA, pH 7.5.
Quality LevelMQ100
Applications
Biological Information
Specific Activity≥10 units/mg protein
Physicochemical Information
Contaminantsβ-N-acetylhexosaminidase, α-fucosidase, β-galactosidase, α- and β-mannosidase, proteases: none detected
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Storage and Shipping Information
Ship Code Blue Ice Only
Toxicity Standard Handling
Storage -20°C
Avoid freeze/thaw Avoid freeze/thaw
Do not freeze Ok to freeze
Special InstructionsFollowing initial thaw, aliquot and freeze (-20°C).
Packaging Information
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Numer katalogowy GTIN
362300-1SET 04055977213669

Documentation

N-Glycosidase F, Chryseobacterium meningosepticum, Recombinant, E. coli MSDS

Title

Safety Data Sheet (SDS) 

N-Glycosidase F, Chryseobacterium meningosepticum, Recombinant, E. coli Certificates of Analysis

TitleLot Number
362300

References

Przegląd literatury
Fan, J.-Q., and Lee, Y.C. 1997. J. Biol. Chem. 272, 27058.
Altmann, F., et al. 1995. Glycoconj. J. 12, 84.
Data Sheet

Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.

Revision19-March-2019 JSW
SynonymsPeptide-N-glycosidase F, Peptide-N⁴-(acetyl-β-glucosaminyl)-asparagine Amidase, PNGase F, Glycopeptidase F
DescriptionRecombinant, Chryseobacterium meningosepticum N-Glycosidase F expressed in E. coli. This enzyme from Chryseobacterium (formerly Flavobacterium) meningosepticum catalyzes the hydrolysis of N-glycans from high mannose, hybrid and complex glycopeptides, and from glycoproteins generating asparagine-linked oligosaccharides. Supplied with a series of buffers.
FormLiquid
FormulationEnzyme supplied in 50 mM NaCl, 20 mM Tris-HCl, 1 mM EDTA, pH 7.5.
Recommended reaction conditions

• Reaction Buffer (Component No. KP31859-1EA): 1 vial, 1 ml, 100 mM sodium phosphate, 0.1% sodium azide, pH 7.5, supplied as a 5X solution
• Denaturation Solution (Component No. KP31860-1EA): 1 vial, 200 µl, 2% SDS, 1 M β-mercaptoethanol
• Detergent Solution (Component No. KP31861-1EA): 1 vial, 200 µl, 15% NP-40 solution
• Tris Reaction Buffer (Component No. KP31865-1EA): 1 vial, 1 ml, 50 mM Tris-HCl, pH 8.0
• N-Glycosidase F, Chryseobacterium meningosepticum, Recombinant, E. coli (Component No. KP31858-100MIU): 1 vial, 100 mU
Note: Tris Reaction Buffer has been included as an alternative reaction buffer because phosphate buffers should be avoided if mass spectrometry is used in downstream analysis.

Procedure for Deglycosylation (Denaturing Conditions)
The amount of enzyme required for deglycosylation depends on the substrate used, the incubation conditions, and the exact application. This protocol is provided only as a guide. Assay conditions should be adjusted as required for individual applications.
1. Prepare 50 to 500 µg glycoprotein solution in 45 µl 1X Reaction. Add 2.5 µl Denaturation Solution.
2. Denature glycoprotein by heating at 100°C for 5 min. Allow mixture to cool.
3. Add 2.5 µl Detergent Solution.
4. Add 2 µl N-Glycosidase to the reaction mixture and incubate for 2 h to overnight at 37°C.
CAS number83534-39-8
EC number3.5.1.52
Contaminantsβ-N-acetylhexosaminidase, α-fucosidase, β-galactosidase, α- and β-mannosidase, proteases: none detected
Specific activity≥10 units/mg protein
Unit definitionOne unit is defined as the amount of enzyme that will completely catalyze the release of N-linked oligosaccharides from 1 µmol denatured ribonuclease in 1 min at 37°C, pH 7.5.
Storage Avoid freeze/thaw
-20°C
Do Not Freeze Ok to freeze
Special InstructionsFollowing initial thaw, aliquot and freeze (-20°C).
Toxicity Standard Handling
ReferencesFan, J.-Q., and Lee, Y.C. 1997. J. Biol. Chem. 272, 27058.
Altmann, F., et al. 1995. Glycoconj. J. 12, 84.