Ciliary Neurotrophic Factor (CNTF)?is a 22 kDa cytokine initially identified for its role in supporting parasympathetic neuron survival. It signals through a tripartite receptor complex to activate JAK-STAT pathways, particularly STAT3. Primarily expressed in Schwann cells and astrocytes, CNTF promotes neuronal survival, differentiation, and regeneration in both CNS and PNS. It demonstrates neuroprotective effects in models of ALS, Huntington's disease, and retinal degeneration. Unlike classical cytokines, CNTF lacks a signal peptide and is released after cellular injury. Current research explores its therapeutic potential for neurodegenerative disorders and metabolic regulation.Besides,Erythropoietin (EPO) has been identified as an interactor of CNTF, thus a functional binding ELISA assay was conducted to detect the interaction of recombinant rat CNTF and recombinant human EPO. Briefly, CNTF was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to EPO-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-CNTF 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. The binding activity of recombinant rat CNTF and recombinant human EPO was shown in Figure 1, and this effect was in a dose dependent manner.