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- Assay Type Competitive ELISA
- Validated Sample Types Tissue Extracts
- Cited Sample Types Supernatant, Homogenized Larva
- Sensitivity 197.8 pg/mL.
- Species 20-Hydroxyecdysone is identical across species
- Assay Duration 2.5 Hours
- Samples/Plate 39 in Duplicate
- Readout Colorimetric, 450 nm
- Standard Curve
The 20-Hydroxyecdysone ELISA Kit quantitatively measures 20-Hydroxyecdysone in tissue extracts. The 20-Hydroxyecdysone ELISA Kit is a competitive ELISA with a run time of 2.5 hours. Please read the complete kit insert for more information before performing this assay.
Use our provided 20-Hydroxyecdysone standard to generate a standard curve for the assay. Pipette the standards or diluted samples into a transparent microtiter plate coated with our goat anti-rabbit IgG antibody. Add the 20-Hydroxyecdysone peroxidase conjugate and the 20-Hydroxyecdysone polyclonal rabbit antibody. Then incubate the mixture covered at room temperature, shaking for 2 hours. The immunological reaction occurs between the anti-20-Hydroxyecdysone antibody, the 20-Hydroxyecdysone antigen in the sample or standard, and the 20-Hydroxyecdysone-peroxidase conjugate. As the 20-Hydroxyecdysone concentration in the sample increases, the bound 20-Hydroxyecdysone-peroxidase conjugate decreases, causing a decrease in signal and vice versa.
After the 2-hour incubation, wash away the excess 20-Hydroxyecdysone-peroxidase conjugate and add the TMB substrate. The TMB substrate reacts with the bound 20-Hydroxyecdysone-peroxidase conjugate generating a signal detected by a plate reader at 450nm. Use the intensity and the standard curve to calculate the 20-Hydroxyecdysone concentration in the samples.
The first insect molting hormone, Ecdysone, was isolated from silkworm pupae in 1954. Later in 1996, 20-hydroxyecdysone was identified and recognized as a derivative of Ecdysone. Ecdysone and its derivatives were identified as a group of steroid hormones that regulate metamorphosis, cell death, and reproduction in arthropods and as a widely distributed steroid in plant species, spanning around one million species. During metamorphosis, the level of 20-hydroxyecdysone in the insect body changes and activates signaling through an ecdysone receptor to mature the larval and/or chrysalis into an adult insect. 20-hydroxyecdyone hormone in plants has also gained interest because of the potential role of the hormone in facilitating the defense mechanisms of plants against insects.
Recent studies have also discovered the ability of 20-hydroxyecdysone to increase osteogenesis and bone mass by reducing cartilage degradation and increasing protein synthesis in humans. Thus 20-hydroxyecdysone has recently gained medical research interest in the treatment of osteoporosis and marketing interest for bodybuilding supplements to increase muscle mass.