GLS (EC3.5.1.2) is mainly found in higher animals and some bacteria and plant roots, catalyzing the
hydrolysis of glutamine into glutamic acid and ammonia, which plays an important role in the regulation of
nitrogen metabolism, especially the regulation of free ammonia and urea metabolism.
The kit uses the indophenol blue colorimetric method to determine ammonia produced by glutamine of
GLS-catalyzed to indicate activity
Reagents and Equipment Required but Not Provided:
Spectrophotometer, adjustable pipette, mortar/homogenizer, centrifuge, 1 mL glass cuvette, ice and
distilled water.
Sample preparation:
1. Tissues: The mass (g): volume of distilled water(mL)= 1:5-10, suggested 0.1g of tissues, add 1 mL of
Extract solution and fully grind. Centrifuge at 12000g at 4 °C for 15 min, then take supernatant on ice
to be tested.
2. Bacteria or cells
Accordance ratio bacteria or cell amount (104): volume of Extract solution (mL)=500~1000:1.
Suggested 5 million with 1 mL of Extract solution. Use ultrasonic to splitting bacteria or cell (placed
on ice, powder: 300W, work time 3s，interval 7s，total time 3 min). Centrifuge at 12000g at 4°C for
15 min. then take supernatant on ice to be tested.
