Dermokine (DMKN), a 18-kDa secreted protein encoded by the?DMKN?gene in the epidermal differentiation complex, is primarily expressed in terminally differentiating keratinocytes of the skin epidermis. Functioning as a matricellular protein, DMKN regulates epidermal barrier formation and inflammation by modulating cell adhesion, cytokine signaling , and cornified envelope maturation. It localizes to the granular layer and stratum corneum, where it interacts with structural proteinsand lipids to enhance skin integrity. Dysregulation of DMKN is linked to inflammatory skin disorders and wound healing defects, highlighting its role in epidermal homeostasis and host defense.Besides,Suprabasin (SBSN) has been identified as an interactor of DMKN, thus a functional binding ELISA assay was conducted to detect the interaction of recombinant human DMKN and recombinant human SBSN. Briefly, DMKN was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to SBSN-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-DMKN 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 human DMKN and recombinant human SBSN was shown in Figure 1, the EC50?for this effect is 0.54ug/mL.