Active Aldolase B, Fructose Bisphosphate (ALDOB)

Aldolase B, Fructose Bisphosphate (ALDOB, encoded by the ALDOB gene) is a key glycolytic enzyme primarily expressed in the liver, kidney, and small intestine. It catalyzes the cleavage of fructose-1,6-bisphosphate and fructose-1-phosphate, critical for fructose metabolism. Deficiencies in ALDOB cause hereditary fructose intolerance (HFI), an autosomal recessive disorder leading to fructose-induced hypoglycemia, liver damage, and other complications. As a vital metabolic regulator, ALDOB maintains energy homeostasis by linking fructose catabolism to glycolytic and gluconeogenic pathways. ALDOB interacts with GAPDH, a housekeeping enzyme, likely facilitating metabolic pathway crosstalk and efficient energy metabolism coordination.Thus a functional ELISA assay was conducted to detect the interaction of recombinant human ALDOB and recombinant mouse GADPH. Briefly, ALDOB was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to GADPH-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-ALDOB 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 ALDOB and recombinant mouse GADPH was shown in Figure 1, the EC50 for this effect is 0.04916µg/mL.