Acyl Coenzyme A Synthetase Long Chain Family, Member 6 (ACSL6) is an enzyme belonging to the acyl - coenzyme A synthetase long - chain family. It activates long - chain fatty acids by catalyzing their conversion to acyl - CoA esters. This process is crucial for fatty acid metabolism, as acyl - CoA esters are key intermediates in various metabolic pathways, such as beta - oxidation and lipid synthesis. ACSL6 is mainly expressed in certain tissues like the liver and adipose tissue, playing a significant role in maintaining lipid homeostasis. Dysregulation of ACSL6 has been associated with several metabolic disorders, including obesity and diabetes.ACSL6 activates fatty acids to acyl - CoA, which is a substrate for CPT1A. Their interaction is essential for the transport of fatty acids into the mitochondria for beta - oxidation.thus a functional binding ELISA assay was conducted to detect the interaction of recombinant mouse ACSL6 and recombinant human CPT1A .Briefly, biotin-linked ACSL6 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100μl were then transferred to CPT1A-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST 3 times and incubation with Streptavidin-HRP for 30min, then wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50μl stop solution to the wells and read at 450nm immediately. The binding activity of recombinant mouse ACSL6 and recombinant human CPT1A was shown in Figure 1, the EC50?for this effect is 0.289ug/mL.