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484400
Sigma-AldrichNitrocefin - CAS 41906-86-9 - Calbiochem
A chromogenic β-lactamase substrate that undergoes distinctive color change from yellow as the amide bond in the β-lactam ring is hydrolyzed by β-lactamase.
More>>A chromogenic β-lactamase substrate that undergoes distinctive color change from yellow as the amide bond in the β-lactam ring is hydrolyzed by β-lactamase. Less<<
Nitrocefin - CAS 41906-86-9 - Calbiochem MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
A 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.
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.
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Storage
-20°C
Protect from Light
Protect from light
Do not freeze
Ok to freeze
Special Instructions
Following reconstitution, aliquot and freeze (-20°C). Stock solution may be stored at -20°C for up to 2 weeks.
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.
数据表
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.
A 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.
Form
Orange-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 number
41906-86-9
Chemical formula
C₂₁H₁₆N₄O₈S₂
Structure formula
Purity
≥95% by UV
Solubility
Preparing 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
Protect from light
-20°C
Do Not Freeze
Ok to freeze
Special Instructions
Following reconstitution, aliquot and freeze (-20°C). Stock solution may be stored at -20°C for up to 2 weeks.
Toxicity
Flammable
References
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.
