484400 Nitrocefin - CAS 41906-86-9 - Calbiochem

484400
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484400-5MG
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      Glass bottle 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 SDS

      Title

      Safety Data Sheet (SDS) 

      Nitrocefin - CAS 41906-86-9 - Calbiochem Certificates of Analysis

      TitleLot Number
      484400

      References

      Reference overview
      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.
      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.

      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.