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Active Glutamine synthetase (GS)

GLNS; GLUL; Glutamate-Ammonia Ligase; Glutamate decarboxylase

ACTIVITY TEST

Glutamine synthetase (GS), encoded by the GLUL gene, is a key enzyme in nitrogen metabolism that catalyzes the ATP-dependent conversion of glutamate and ammonia into glutamine. Widely expressed in tissues like the liver, brain, and kidneys, it maintains nitrogen homeostasis—detoxifying ammonia in the liver and supporting neurotransmitter synthesis in the brain. GS also plays a critical role in cancer metabolism, as rapidly dividing tumor cells rely on glutamine for energy and biosynthesis. GS and GLS (glutaminase) function in a cyclic relationship: GLS breaks down glutamine into glutamate, which GS then uses to resynthesize glutamine, though they do not directly bind physically.To detect the activity of recombinant GS , a functional ELISA assay was performed to evaluate the interaction between recombinant human GS and recombinant human GLS.Briefly, GLS was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to GS-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-GLS pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody for 1h at 37℃, wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50 µL stop solution to the wells and read at 450/630nm immediately.Measured by its binding ability in a functional ELISA. When Recombinant GS is lmmobilized at 2 µg/mL(100 uLwell), the concentration of GLS that produces 50% optimal bindingresponse is found to be approximately 0.00494 µg/mL.

USAGE

Reconstitute in 10mM PBS (pH7.4) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

STORAGE

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

STABILITY

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

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Magazine Citations
The FASEB Journal Cargo proteins of plasma astrocyte-derived exosomes in Alzheimer’s disease pubmed:27511944
Annals of Neurology  High complement levels in astrocyte‐derived exosomes of Alzheimer disease Pubmed:29406582
Journal of Neuroscience Sex differences in the glutamate signaling pathway in juvenile rats Pubmed:28861894
FASEB Journal Deficient neurotrophic factors of CSPG4-type neural cell exosomes in Alzheimer disease Pubmed:29924942
Hepatology Research Glutamine synthetase promotes tumor invasion in hepatocellular carcinoma through mediating epithelial–mesenchymal transition Pubmed: 31652385
FASEB JOURNAL Traumatic brain injury increases plasma astrocyte‐derived exosome levels of neurotoxic complement proteins Pubmed: 31916313
Translational Psychiatry Decreased mitochondrial electron transport proteins and increased complement mediators in plasma neural-derived exosomes of early psychosis Pubmed: 33106473
Drug Repurposing of Asparaginase and Vitamin C Targeting Glutamine Synthetase Improves Anticancer Effect in Metastatic Castration-resistant Prostate …
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