General description

A chromogenic β-lactamase substrate that undergoes distinctive color change from yellow (λ_max = 390 nm at pH 7.0) to red (λ_max = 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.

Legal Information

CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany

Other Notes

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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Packaging

5 mg in Glass bottle

Packaged under inert gas

Preparation Note

Preparing a Nitrocefin (500 µg/ml) Solutionr> Dissolve 1 mg Nitrocefin in 100 µl dimethylsulfoxide (DMSO) and vortex. r> Add 1.9 ml phosphate buffer (100 mM, pH 7) to produce 2 ml total volume.r> This yields a working Nitrocefin solution of 500 µg/ml (~1 mM), which is suitable for most applications.r> Nitrocefin, particularly in solution, is very sensitive to light.

Reconstitution

Following reconstitution, aliquot and freeze (-20°C). Stock solution may be stored at -20°C for up to 2 weeks.

Warning

Toxicity: Flammable (J)