These assays utilize two distinct ELISA formats—Competitive and Sandwich—to provide the most accurate quantification for each specific molecule type.

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Competitive ELISA

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Targets: Insulin, Amylin, Glucagon, and GLP-1

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In this format, the plate is pre-coated with a hormone-protein conjugate. When samples are added, free hormones in the sample compete with the immobilized hormones for binding sites on a biotin-labeled primary antibody.

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• Detection: Bound biotinylated antibodies are captured by Streptavidin-HRP.

• Signal: Following the addition of TMB substrate and Stop Solution, absorbance is measured at 450 nm.

• Interpretation: The signal is inversely proportional to the hormone concentration; higher levels of free hormone in the sample result in lower absorbance readings.

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Sandwich ELISA

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Targets: Leptin, Adiponectin, IL-6, and TNFα

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This format uses a matched antibody pair to "sandwich" the target peptide. The sample is added to a plate pre-coated with a specific capture antibody.

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• Detection: After washing, a biotin-labeled detection antibody is added, followed by Streptavidin-HRP to form a stable detection complex.

• Signal: TMB substrate is added to produce a blue color, which turns yellow upon the addition of Stop Solution.

• Interpretation: The intensity of the yellow color (450 nm) is directly proportional to the concentration of the target peptide in the sample.

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