Human Angiogenesis ELISA Strip II for profiling 8 Cytokines

Angiogenesis shifted from the avascular to vascular states is a key event for sustained tumor growth and cancer progression (1). Angiogenesis as a biological switch process is governed by numerous pro- and anti-angiogenic factors, including vascular endothelial growth factor (VEGF), basic fibroblast growth factor (FGFb), epidermal growth factor (EGF), and transforming growth factor-beta (TGF-b). The mechanism of action of each of these factors is different, as are their origin and the stimuli for their production. The angiogenic switch refers to the balance between pro- and anti- angiogenic factors. Therefore, profiling of these factors is critical to understanding angiogenesis. Signosis’ Angiogenesis ELISA Strip Profiling Assay allows simultaneously profiling 8 angiogenesis cytokines; PDGF-BB, PIGF-1, NGF, SCF, MCP-1, MIP-1a, IL-2, and IL-4. Each well of the strip is coated with a primary antibody against a specific angiogenesis protein and total 8 wells of a strip target 8 different proteins. The difference of these proteins between two samples can be determined through data comparison.

A. Benefits:

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

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

• Quantitative measurment of differences - Unlike membrane-based array assays, the differences of individual proteins among samples can be quantitatively compared. In addition, EA-1042 provides protein standards 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 angiogenesis 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 angiogenesis 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 the angiogenesis cytokines are directly proportional to the color intensity of the test sample. Absorbance is measured spectrophotometrically at 450 nm.

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