GAPDH has a molecular weight of 146 kDa and consists of four subunits, each ranging from 30 to 40 kDa. As a key enzyme in the glycolytic pathway, its primary function is to catalyze the oxidation (dehydrogenation) and phosphorylation of glyceraldehyde-3-phosphate to produce 1,3-bisphosphoglycerate. Due to its widespread distribution across various tissues, consistently high expression levels, and stable expression unaffected by external inducers, GAPDH is widely used as a standard internal reference in experiments such as qPCR and Western blot (WB). The Anti-GAPDH Antibody-HRP is produced by conjugating horseradish peroxidase (HRP) to a GAPDH antibody through a specific process. During WB experiments, the addition of luminol (Solution A) and hydrogen peroxide (Solution B) initiates a chemiluminescent reaction. In the presence of HRP, luminol reacts with hydrogen peroxide to form an unstable peroxide intermediate, which rapidly decomposes into an electronically excited intermediate. As this intermediate returns to its ground state, light is emitted, enabling the detection of the GAPDH reference protein.