Succinate Dehydrogenase Complex Subunit A (SDHA) is an integral protein of the succinate dehydrogenase complex, also known as Complex II in the mitochondrial respiratory chain. It has the active site with flavin adenine dinucleotide (FAD) that catalyzes the oxidation of succinate to fumarate, playing a key role in the tricarboxylic acid (TCA) cycle and electron transport, which contributes to ATP production for cellular energy needs.SDHA binds to Succinate Dehydrogenase Complex Subunit B (SDHB) to form a stable structure, ensuring the proper assembly and function of Complex II for efficient electron transfer and metabolic reactions.Thus a functional ELISA assay was conducted to detect the interaction of recombinant mouse SDHA and recombinant mouse SDHB.
Briefly, SDHA was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to SDHB-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-SDHA 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 mouse SDHA and recombinant mouse SDHB was shown in Figure 1, and this effect was in a dose dependent manner.