Troponin T Type 1, Slow Skeletal (TNNT1) is a protein predominantly expressed in slow-twitch skeletal muscles. It is a crucial component of the troponin complex, which plays a vital role in the regulation of muscle contraction. TNNT1 helps to anchor the troponin complex to tropomyosin on the thin filaments of muscle cells. Mutations in the TNNT1 gene can lead to various myopathies, disrupting normal muscle function. It has a specific structure and function that enables it to interact precisely with other troponin subunits.Besides,it has been identified that interaction between TNNT1 and TNNC2 contributes to allowing TNNC2's calcium-induced change to be relayed, triggering muscle contraction regulation.Thus, a functional binding ELISA assay was used to detect the biological activity of TNNT1Briefly, TNNC2 was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to TNNT1-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-TNNC2 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 TNNT1 is lmmobilized at 2 ug/mL(100 uLwell), the concentration of TNNC2 that produces 50% optimal bindingresponse is found to be approximately 0.00597 ug/mL.