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If you prefer an end‑point assay, skip the kinetic read and incubate the plate for a fixed time (e.g., 10 min) before adding stop solution and measuring a single absorbance value. Zoikhem Lab Choye BEST
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| Category | Item | Details / Recommended Brand | |----------|------|-----------------------------| | | Choye‑A (substrate) | 10 mM stock in 50 % DMSO, store @ –20 °C, protect from light | | | Choye‑B (product standard) | 1 mM in water, used for calibration curve | | | Reaction Buffer | 50 mM Tris‑HCl, pH 7.5, 150 mM NaCl, 5 mM MgCl₂ | | | Stop Solution | 0.5 M HCl (prepared fresh) | | | Optional Inhibitor (for controls) | e.g., PMSF 1 mM (if serine protease) | | Enzyme Samples | Crude lysate or purified enzyme | Adjust to 0.1–1 mg mL⁻¹ protein (Bradford assay) | | Plates & Consumables | 96‑well clear flat‑bottom microplate (UV‑transparent) | Polystyrene, low‑binding preferred | | | Multichannel pipette (8‑ or 12‑channel) | 10 µL–200 µL range | | | Sealers (adhesive film) | Optional, to prevent evaporation | | Instrumentation | Plate reader (spectrophotometer) | Capable of 620 nm absorbance, kinetic mode (30 s intervals) | | | Temperature‑controlled incubator or plate reader with built‑in temp control | Set to 30 °C (or assay‑specific temperature) | | Miscellaneous | Ice bucket, vortex mixer, centrifuge (micro‑centrifuge) | – |
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