The Cortisol, BioAssay™ ELISA Kit is a competitive ELISA intended for the quantitative determination of Cortisol in fish serum, plasma and tissue homogenates.||Detection Range:|0.0023-10ng/ml||Sensitivity:|<0.0023ng/ml||Precision: |Intra-assay Precision: CV <8% |Inter-assay Precision: CV <10%||Specificity:|This assay has high sensitivity and excellent specificity for detection of fish Cortisol. No significant cross-reactivity or interference between fish Cortisol and analogues was observed.|Note: Limited by current skills and knowledge, it is impossible for us to complete the crossreactivity detection between fish Cortisol and all the analogs. Therefore, crossreaction may still exist.||Kit Components:|227575A: Microtiter Strips, 1x48 wells or 1x96 wells. Pre-coated; ready to use. |227575B: Standard, 1x1vial or 2x1vial|227575C: Cortisol, 100X, 1x30ul or 1x60ul|227575D: HRP, 100X, 1x60ul or 1x120ul|227575E: Biotin Diluent, 1x5ml or 1x10ml|227575F: HRP Diluent, 1x10ml or 1x20ml|227575G: Sample Diluent, 2x20ml|227575H: Wash Buffer, 25X, 1x10ml or 1x20ml|227575J: TMB Substrate, 1x5ml or 1x10ml|227575K: Stop Solution, 1x5ml or 1x10ml||Precaution:|The Stop Solution (227575K) included for use in this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.||Storage and Stability:|Store all kit components at 4°C. Stable for 6 months after receipt. Once opened, the kit is stable for 1 month providing this is within the expiration date of the kit. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.||Assay Summary:|1. Prepare reagents and samples as instructed.|2. Set a Blank well without any solution|3. Add 50ul Standard or sample to each well.|4. Add 50ul Cortisol to each well (Not to the Blank well).|5. Incubate for 40 minutes at 37°C.|6. Aspirate and wash 3 times.|7. Add 100ul HRP to each well (Not to the Blank well).|8. Incubate for 30 minutes at 37°C. |9. Aspirate and wash 5 times,|10. Add 90ul TMB Substrate to each well. |11. Incubate for 20 minutes at 37°C. Protect from light|12. Add 50ul Stop Solution to each well. Read at 450nm within 5 minutes.