Larvae were washed of treatment media and pooled (n=30), then snap frozen in liquid nitrogen. Adult zebrafish were euthanized in tricaine and snap frozen in liquid nitrogen and frozen tissues were minced with a razor. Samples were homogenized in PBS using an automatic pestle grinder. A liquid-liquid extraction was performed with ethyl acetate and the organic, cortisol-containing phase was collected. The solvent was evaporated under a stream of nitrogen gas, and the cortisol extract was re-dissolved in Cortisol ELISA Extraction Buffer (Neogen Corporation). Samples were assayed for cortisol content using the Neogen Cortisol ELISA kit and quantified with a standard curve.
Cortisol elisa extraction buffer
Manufactured by Neogen
The Cortisol ELISA Extraction Buffer is a laboratory reagent used in the extraction and preparation of samples for cortisol enzyme-linked immunosorbent assay (ELISA) testing. It is designed to facilitate the extraction of cortisol from various biological matrices, such as serum, plasma, or saliva, prior to analysis.
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Lab products found in correlation
2 protocols using cortisol elisa extraction buffer
1
Quantifying Cortisol in Zebrafish Tissues
2
Cortisol Quantification in Zebrafish
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Zebrafish embryos were washed with the larval medium and snap-frozen in liquid nitrogen. Embryos were then homogenized in 1 ml of PBS using an automatic pestle grinder. Each sample was extracted twice with 5 ml of diethyl ether. Cortisol containing organic phase was collected and evaporated. The extracts were dissolved in Cortisol ELISA Extraction Buffer (Neogen) and were assayed using Neogen Cortisol ELISA kit according to the manufacture's protocol.
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