Glutamate Receptor, Ionotropic, AMPA 1 (GRIA1) is a key subunit of AMPA receptors that mediates fast excitatory synaptic transmission in the brain. It forms heterotetrameric complexes with other subunits like GRIA2, which critically regulates receptor function. While GRIA1-containing receptors exhibit calcium permeability, co-assembly with GRIA2 reduces calcium influx and stabilizes receptor expression at synapses. This GRIA1-GRIA2 interaction ensures balanced synaptic plasticity, underlying learning and memory processes. Dysfunction of this partnership contributes to neurological disorders including epilepsy and cognitive impairment. Thus a functional binding ELISA assay was conducted to detect the interaction of recombinant human GRIA1 and recombinant human GRIA2 . Briefly, biotin-linked GRIA1 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100μl were then transferred to GRIA2-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST 3 times and incubation with Streptavidin-HRP for 30min, then wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50µl stop solution to the wells and read at 450nm immediately. The binding activity of recombinant human GRIA1 and recombinant human GRIA2 was shown in Figure 1, the EC50 for this effect is 0.116ug/mL.