Radioimmunoassay for the detection of leptin in human serum.

摘要: Human leptin, which is encoded by the obese (ob) gene, secreted specifically from adipocytes and involved in regulation of satiety energy consumption. We developed a radioimmunoassay for determination leptin human serum using polyclonal antibodies generated rabbits against C-terminal fragment leptin(126-140), coupled to hemocyanin. The sensitivity assay was app. 5 pmol/l leptin(126-140) equivalent 0.5 fmol/tube. intra-assay variation at 100 less than 4.8% interassay 8.3%. Dilution curves samples containing high levels were parallel standard curve. Following G-50 Sephadex chromatography single specific peak detected 16 kd. procedure compared well commercially available (Linco, St. Louis, USA) directed intact recombinant protein (R = 0.96; p < 0.0001). Serum significantly higher plasma (app.20%) over wide range Levels leptin126-140 immunoreactivity not altered meals no day-to-day found. In group 148 healthy female 108 male subjects with BMI between 18.2 40 kg/m2 there significant difference sexes circulating when tested identical (p 0.001). both positively related 0.001) analysed lean whereas this relation apparent. No age detectable up 30 kg/m2. These data support an important role body fat stores modulated gender factors.