GLS (EC3.5.1.2) is mainly found in higher animals, some bacteria and plant roots, catalyze 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/microplate reader, adjustable pipette, balance, mortar/homogenizer, centrifuge,
micro glass cuvette/96 well flat-bottom plate, ice and distilled water.
Sample preparation:
1. Tissue: The mass (g): Extract solution volume (mL)= 1:5-10, suggested 0.1g of tissue, add 1 mL of
Extract solution. After homogenizing on ice, centrifuge 12000g at 4 ° C for 15 min, then take
supernatant 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.
