484400 Nitrocefin - CAS 41906-86-9 - Calbiochem

484400
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      Référence DisponibilitéConditionnement Qté Prix Quantité
      484400-5MG
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      Disponibilité limitéeDisponibilité limitée
      En stock 
      Interrompu(e)
      Disponible en quantités limitées
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          Flacon en verre 5 mg
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          Description
          OverviewA chromogenic β-lactamase substrate that undergoes distinctive color change from yellow (λmax = 390 nm at pH 7.0) to red (λmax = 486 nm at pH 7.0) as the amide bond in the β-lactam ring is hydrolyzed by β-lactamase. Nitrocefin is sensitive to hydrolysis by all known lactamases produced by Gram-positive and Gram-negative bacteria. Also useful for the detection of β-lactamase patterns from bacterial cell extracts by isoelectric focusing. Has been used in competitive inhibition studies in developmental work on β-lactamase-resistant antibiotics.
          Catalogue Number484400
          Brand Family Calbiochem®
          Synonyms3-(2,4-Dinitrostyryl)-(6R, 7R)-7-(2-thienylacetamido)-ceph-3-em-4-carboxylic Acid, E-isomer
          References
          ReferencesGuay, R., et al. 1980. IRCS Med. Science 8, 209.
          King, A., et al. 1980. Antimicrob. Agents Chemother. 17, 165.
          Matthew, M., et al. 1975. J. Gen. Microbiol. 88, 169.
          O’Callaghan, C.H., et al. 1972. Antimicrob. Agents Chemother. 1, 283.
          Product Information
          CAS number41906-86-9
          FormOrange-yellow solid
          Hill FormulaC₂₁H₁₆N₄O₈S₂
          Chemical formulaC₂₁H₁₆N₄O₈S₂
          Structure formula ImageStructure formula Image
          Quality LevelMQ200
          Applications
          Biological Information
          Purity≥95% by UV
          Physicochemical Information
          Dimensions
          Materials Information
          Toxicological Information
          Safety Information according to GHS
          Safety Information
          R PhraseR: 10

          Flammable.
          S PhraseS: 16

          Keep away from sources of ignition - No Smoking.
          Product Usage Statements
          Storage and Shipping Information
          Ship Code Ambient Temperature Only
          Toxicity Flammable
          Hazardous Materials Attention: Due to the nature of the Hazardous Materials in this shipment, additional shipping charges may be applied to your order. Certain sizes may be exempt from the additional hazardous materials shipping charges. Please contact your local sales office for more information regarding these charges.
          Storage -20°C
          Protect from Light Protect from light
          Do not freeze Ok to freeze
          Special InstructionsFollowing reconstitution, aliquot and freeze (-20°C). Stock solution may be stored at -20°C for up to 2 weeks.
          Packaging Information
          Packaged under inert gas Packaged under inert gas
          Transport Information
          Supplemental Information
          Specifications

          Documentation

          Nitrocefin - CAS 41906-86-9 - Calbiochem FDS

          Titre

          Fiche de données de sécurité des matériaux (FDS) 

          Nitrocefin - CAS 41906-86-9 - Calbiochem Certificats d'analyse

          TitreNuméro de lot
          484400

          Références bibliographiques

          Aperçu de la référence bibliographique
          Guay, R., et al. 1980. IRCS Med. Science 8, 209.
          King, A., et al. 1980. Antimicrob. Agents Chemother. 17, 165.
          Matthew, M., et al. 1975. J. Gen. Microbiol. 88, 169.
          O’Callaghan, C.H., et al. 1972. Antimicrob. Agents Chemother. 1, 283.
          Fiche technique

          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.

          Revision13-May-2008 JSW
          Synonyms3-(2,4-Dinitrostyryl)-(6R, 7R)-7-(2-thienylacetamido)-ceph-3-em-4-carboxylic Acid, E-isomer
          DescriptionA chromogenic β-lactamase substrate that undergoes distinctive color change from yellow (λmax = 390 nm at pH 7.0) to red (λmax = 486 nm at pH 7.0) as the amide bond in the β-lactam ring is hydrolyzed by β-lactamase. Nitrocefin is sensitive to hydrolysis by all known lactamases produced by Gram-positive and Gram-negative bacteria. Also useful for the detection of β-lactamase patterns from bacterial cell extracts by isoelectric focusing. Has been used in competitive inhibition studies in developmental work on β-lactamase-resistant antibiotics.
          FormOrange-yellow solid
          Intert gas (Yes/No) Packaged under inert gas
          Recommended reaction conditions
          Techniques for the Rapid Detection of β-Lactamase Using Nitrocefin 1. Direct Plate Method Add one drop of the Nitrocefin working solution on to the surface of the colony. If the isolate is a high β-lactamase producer then the colony and the surrounding area will turn red quickly. 2. Slide Method Add one drop of the Nitrocefin working solution on to the surface of a clean glass slide. Using a sterile loop, pick one colony from the plate and emulsify into the Nitrocefin drop. Report as positive for β-lactamase if the color changes from yellow to red within 30 min. NOTE: Protect the slide from desiccation during the waiting period. 3. Broth Method Add four drops of the Nitrocefin solution to 1 ml of the grown culture. Report as positive for β-lactamase if the color changes to red within 30 min. 4. Broken Cell Method Sonicate 1 ml of the culture in order to break open the cells. Add 4 drops of the Nitrocefin working solution. Report as positive for β-lactamase if the color changes to red within 30 min. 5. Paper Disc Spot Method Place a Whatman No. 1 filter paper disc (diameter 7 cm) in a petri dish and impregnate with 5 ml of the Nitrocefin working solution. Apply an isolated colony to the impregnated paper disc using a sterile loop. A pink to red reaction developing within 15 min indicates the presence of β-lactamase. NOTE: The impregnated paper disc is stable for one day, if protected from light to avoid degradation. 6. Spectrophotometric Assays for Determining β-Lactamase Activity The working solution of Nitrocefin (500 µg/ml) is diluted ten-fold in buffer (0.1 M phosphate; 1 mM EDTA, pH 7.0). Spectrophotometric assays for β-Lactamase using Nitrocefin are carried out by measuring changes in absorbance at 486 nm. The molar extinction coefficient of hydrolyzed Nitrocefin at 486 nm is 20,500 M-1 cm-1. Test samples of the finished product for performance with control cultures.
          CAS number41906-86-9
          Chemical formulaC₂₁H₁₆N₄O₈S₂
          Structure formulaStructure formula
          Purity≥95% by UV
          SolubilityPreparing a Nitrocefin (500 µg/ml) Solution
          • Dissolve 1 mg Nitrocefin in 100 µl dimethylsulfoxide (DMSO) and vortex.
          • Add 1.9 ml phosphate buffer (100 mM, pH 7) to produce 2 ml total volume.
          • This yields a working Nitrocefin solution of 500 µg/ml (~1 mM), which is suitable for most applications.
          • Nitrocefin, particularly in solution, is very sensitive to light.
          Storage -20°C
          Protect from light
          Do Not Freeze Ok to freeze
          Special InstructionsFollowing reconstitution, aliquot and freeze (-20°C). Stock solution may be stored at -20°C for up to 2 weeks.
          Toxicity Flammable
          ReferencesGuay, R., et al. 1980. IRCS Med. Science 8, 209.
          King, A., et al. 1980. Antimicrob. Agents Chemother. 17, 165.
          Matthew, M., et al. 1975. J. Gen. Microbiol. 88, 169.
          O’Callaghan, C.H., et al. 1972. Antimicrob. Agents Chemother. 1, 283.