Mouse Obesity ELISA Strip for Profiling 8 Cytokines

Obesity increases the risk for the metabolic syndrome, diabetes, hypertension, atherosclerosis, and thrombosis. A number of proteins have been identified to be relevant to the development of the metabolic syndrome, diabetes, and cardiovascular disease with obesity. These include adiponectin, leptin, TNFa, IGF-1, Resistin, TGFß, IL-6, and PAI-1. Plasma concentrations of these proteins are usually measured by ELISA. To systematically examine the effects, Signosis developed an ELISA strip which allows simultaneous determination of 8 proteins: leptin, TNFa, IGF-1, IL-6, VEGF, IL-1a, IL-1b, and MCP-1. The difference of these proteins between two samples can be determined through data comparison. Therefore, it facilitates the discovery of the change of these proteins in different samples.

A. Benefits:

• Multiplex - A single ELISA strip allows the measurement of 8 obesity cytokines.

• Flexible - The comparison can be made from 2 samples to 12 samples with a single plate.

• Quantitative measurement of differences - Unlike membrane-based array assays, the differences of individual proteins among samples can be quantitatively compared. In addition, EA-1072 provides individual proteins for making standard curves.

• No need of capital equipment - Unlike beads-based assays, no capital equipment is required.

• Simple procedure -All in one system with a pre-coated 12 X 8 well plate.

B. Principle of the technology

In each well of the strip, a primary antibody against a specific obesity cytokine is coated and 8 wells of the strip are coated with 8 different antibodies. Therefore, total 8 wells of a strip allow measurement of 8 different cytokines. The test sample is allowed to react simultaneously with pairs of two antibodies, resulting in the obesity cytokines being sandwiched between the solid phase and enzyme-linked antibodies. After incubation, the wells are washed to remove unbound-labeled antibodies. A HRP substrate, TMB, is added to result in the development of a blue color. The color development is then stopped with the addition of Stop Solution changing the color to yellow. The concentrations of obesity cytokines are directly proportional to the color intensity of the test sample. Absorbance is measured spectrophotometrically at 450 nm.

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