Troponin C Type 2, Fast (TNNC2) is a protein predominantly expressed in fast - twitch skeletal muscles. It is a crucial component of the troponin complex. TNNC2 binds calcium ions, which initiates a conformational change in the troponin - tropomyosin complex. This change allows myosin to interact with actin, enabling muscle contraction. Mutations in the TNNC2 gene can lead to various skeletal muscle disorders affecting muscle function.It has been reported that the binding of TNNC2 and TNNT3 plays a key role in the process of rapid skeletal muscle contraction.Thus a functional ELISA assay was conducted to detect the interaction of recombinant human TNNC2 and recombinant human TNNT3.
Briefly, TNNC2 was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to TNNT3-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. The binding activity of recombinant human TNNC2 and recombinant human TNNT3 was shown in Figure 1, the EC50 for this effect is 0.003ug/mL.