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Beta-lactamases, especially important in Gram-negative pathogens, open the four-atom beta-lactam ring, which lets bacteria resist penicillins, cephalosporins, cephamycins and carbapenems. Inhibiting these enzymes keeps the antibiotics from being broken down.
Kit vs Kit Plus
The only difference is the enzyme: LAC100K contains buffer, penicillin G and dye but no enzyme, while LAC100KE adds 55 µl of 100 x E. coli beta-lactamase. Both are for 100 assays.
| Kit (enzyme not included), 100 assays (LAC100K) | 100 assays |
|---|---|
| Kit Plus (E. coli beta-lactamase included), 100 assays (LAC100KE) | 100 assays |
Kit Components
| Component | Kit (enzyme not included), 100 assays (LAC100K) | Kit Plus (E. coli beta-lactamase included), 100 assays (LAC100KE) |
|---|---|---|
| 10 x Buffer | 550 µl | 550 µl |
| 50 x Penicillin G | 110 µl | 110 µl |
| 10 x Fluorescence dye | 260 µl | 260 µl |
| 100 x E. coli beta-lactamase | Not included | 55 µl |
The 10 x Buffer is 100 mM HEPES, pH 7.5, 500 mM KCl.
Assay Principle
Beta-lactamase hydrolyzes the penicillin G substrate. A dye binds the hydrolyzed beta-lactam and becomes fluorescent at 535 nm (excitation 485 nm), so the signal rises with enzyme activity and falls with inhibitors.
Specifications
| Assay Target | Beta-lactamase activity |
|---|---|
| Target Type | Beta-Lactamase |
| Organism | E. coli |
| Assay Type | Enzyme Activity Assay; Inhibitor Screening Assay |
| Detection Method | Fluorescence |
| Detection Wavelength | Ex 485 nm / Em 535 nm |
| Plate Format | 384-well or 96-well plate |
| Instrument Required | Fluorescence microplate reader |
| Assay Time | About 65 min (60 min reaction + 5 min detection) |
| Compatibility | Not compatible with samples containing free amino acids or buffers with DTT or EDTA |
| Size | 100 assays |
| Storage | Please contact us to confirm storage conditions. |
| Shipping | Shipped frozen. |
Protocol
The manufacturer's protocol describes the procedure and reaction volumes.
- E. coli Beta Lactamase Assay (PDF, see Documents): Covers catalog # LAC100K, LAC100KE.
Safety & Handling
Can’t find the exact format you need? We can help you source or customize this beta-lactamase activity assay — including larger pack sizes, bulk reagents, bulk enzyme, or a protocol adapted to your samples. Click Talk to a Scientist, email support@biohippo.com, or explore our Research Services. Our team will be in touch shortly.