A β-lactamase Activity Assay is a laboratory test used to measure the activity of β-lactamase enzymes. These enzymes are produced by certain bacteria and can break down β-lactam antibiotics (like penicillin), rendering them ineffective. This assay is crucial for detecting bacterial resistance to β-lactam antibiotics.

Key Components of the β-lactamase Activity Assay:

1. β-lactamase enzyme: The enzyme of interest that catalyzes the hydrolysis of β-lactam antibiotics.
2. Substrate: A β-lactam antibiotic (e.g., penicillin or cephalosporins) that the β-lactamase will degrade. The substrate can be labeled or have a detectable change when broken down.
3. Detection method: Various methods can be used to detect the breakdown of the β-lactam antibiotic:
   • Colorimetric assays: A chromogenic substrate can change color upon hydrolysis.
   • Fluorometric assays: Fluorescent substrates can emit light when cleaved by the enzyme.
   • Spectrophotometric assays: Monitoring the absorbance of a substance in solution as it is hydrolyzed.
4. Reaction conditions: The assay is performed under controlled conditions, usually at a specific pH and temperature, to optimize enzyme activity.

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