Invitrogen™ 20-Hydroxyecdysone ELISA Kit

ELISA

The 20-Hydroxyecdysone Competitive ELISA quantitates 20-Hydroxyecdysone (20E) in serum, plasma, urine, saliva, dried fecal extracts or cell culture medium.∣Principle of the method: The 20-Hydroxyecdysone Competitive ELISA research-use-only kit is designed to quantitatively measure 20-Hydroxyecdysone independent of species. A 20-Hydroxyecdysone standard is provided to generate a standard curve for the assay and all samples are read off a user-generated standard curve. Standards or diluted samples are pipetted into a clear microtiter plate coated with anti-rabbit IgG antibodies. Anti-rabbit 20-Hydroxyecdysone antibodies and 20-Hydroxyecdysone-peroxidase conjugate is added to the wells. The 20-Hydroxyecdysone-peroxidase conjugate and any 20-Hydroxyecdysone in the sample will compete to bind to the anti-rabbit 20-Hydroxyecdysone antibodies. After incubation, the plate is washed and substrate is added. The substrate reacts with the bound 20-Hydroxyecdysone-peroxidase conjugate. After a 30 min incubation, the reaction is stopped and the intensity of the generated color is detected in a microtiter plate reader at 450 nm. The intensity of the generated color corresponds inversely to the amount of 20-Hydroxyecdysone present in the sample.∣Rigorous validation: Each manufactured lot of this ELISA kit is quality tested for criteria such as sensitivity, specificity, precision, and lot-to-lot consistency. See manual for more information on validation.