Asparaginyl tRNA Synthetase (NARS) is a classII aminoacyltRNA synthetase that specifically ligates asparagine to its cognate tRNA, ensuring accurate translation during protein synthesis. Essential for cell survival and proliferation, NARS maintains proteome fidelity and supports normal cellular metabolism. Beyond translation, NARS participates in stress responses, signaling pathways, and immune regulation. Dysregulation is linked to neurodevelopmental disorders, inflammation, and tumor progression. NARS forms a multisynthetase complex with EPRS to stabilize the complex and coordinate efficient protein synthesis.Briefly, EPRS was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to NARS-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-EPRS 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 mouse NARS is lmmobilized at 2 µg/mL(100 µLwell), the concentration of EPRS that produces 50% optimal bindingresponse is found to be approximately 0.046 µg/mL.