Millipore Technical Publications |
A Novel High Molecular Weight (HMW) Adiponectin ELISA: Validation by Size Exclusion Chromatography and Western Blot Analysis | Product Catalogue | |
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| HMW adiponectin of the 3 circulating multimeric iso-forms have been shown to be a better index of insulin sensitivity than total adiponectin using size exclusion and/or immuno-blotting techniques. We have developed a size specific HMW adiponectin ELISA only requiring simple pretreatment of serum and plasma samples. HMW and total adiponectin levels are determined in 108 subjects and in 20 morbidly obese subjects before and after (1, 3, 6 and 12 months) gastric bypass surgery. The ELISA has a concentration range of 3.125 - 200 ng/mL with 0.8 ng/mL sensitivity. Intra- and interassay CV are < 4% and 10%. Linearity of dilution curves are parallel to calibration curves. FPLC profiles of the enzyme treated samples shows recognition of predominantly HMW Adiponectin. HMW Adiponectin by this method closely compares to Western Blot analysis (y= 0.77x-0.15; r = 0.96, n=56). BMI and sex related changes are more pronounced and significant for HMW and % HMW compared to total Adiponectin. Gastric bypass surgery causes increases in HMW and total Adiponectin, but changes in HMW Adiponectin are more pronounced and precede to the changes in total Adiponectin. This novel HMW adiponectin ELISA after enzyme treatment recognizes predominantly HMW adiponectin, closely compares with Western Blot data, and magnifies BMI and gender associated differences than those for total adiponectin. The new ELISA is analytically robust, simple and rapid and can easily be adopted for high throughput analysis. Click PDF above to view entire poster. |