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

362300
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      Overview

      Replacement Information

      Key Specifications Table

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      Catalog NumberAvailability Packaging Qty/Pack Price Quantity
      362300-1SET
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          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.
          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

          Documentation

          SDS

          Title

          Safety Data Sheet (SDS) 

          Certificates of Analysis

          TitleLot Number
          362300

          References

          Reference overview
          Fan, J.-Q., and Lee, Y.C. 1997. J. Biol. Chem. 272, 27058.
          Altmann, F., et al. 1995. Glycoconj. J. 12, 84.

          Brochure

          Title
          Proteases Technical Bulletin
          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-May-2010 RFH
          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. KP31858-1MIU): 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 -20°C
          Avoid freeze/thaw
          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.