Troponin C Type 1, Slow (TNNC1) is a calcium - binding protein crucial for muscle contraction. Found predominantly in slow - twitch skeletal muscles, it binds calcium ions, triggering conformational changes. This activation enables its interaction with other troponin complex components. TNNC1 strongly binds to Troponin I Type 2, Fast Skeletal (TNNI2), forming a key regulatory unit that controls muscle contraction and relaxation.
Thus a functional ELISA assay was conducted to detect the interaction of recombinant human TNNC1 and recombinant human TNNI2. Briefly, TNNC1 was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to TNNI2-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-TNNC1 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 TNNC1 and recombinant human TNNI2 was shown in Figure 1, the EC50 for this effect is 0.028ug/mL.