Table 1 Comparison of the different steps of the two techniques
From: Optimization of the arginase activity assay micromethod for macrophages and sera
Micromethod | Optimized micromethod | |
|---|---|---|
Equipment | Waterbath, Oven, Incubator and Spectrophotometer | Thermocycler and Spectrophotometer |
Activation | Sample volume: 50 µL Buffer : Tris Waterbath : 56 °C for 15 min Eppendorf tube | Sample volume: 6 µl Buffer: HEPES Thermocycler : 56 °C for 15 min Microplate well |
Reaction | Substrate: Arginine in PBS Proofer at 37 °C for 1 h 30 Addition of 400 µL acidic mix Eppendorf tube | Substrate: Arginine in deionized water Thermocycler à 37 °C for 1 h 30 Addition of 200 µL acidic mix Same Microplate well |
Revelation | Addition of 25 µL ISPF Oven at 100 °C for 45 min Glass tube | Addition of 15 µL ISPF Thermocycler at 96 °C for 45 min Same Microplate same well |
Reading | Spectrophotometer 550 nm Microplate well | Spectrophotometer 550 nm Same Microplate well |