Thyroid Stimulating Hormone, Human, Normal (TSH) BioAssay(TM) ELISA Kit

Intended Use: This Human TSH ELISA Kit is to be used for the in vitro quantitative determination of human thyroid stimulating hormone (TSH) concentrations in serum. It is intended for research use only and not for use in diagnostic procedures. Principle of the Assay: This TSH enzyme linked immunosorbent assay (ELISA) applies a technique called a quantitative sandwich immunoassay. The microtiter plate provided in this kit has been pre-coated with a monoclonal antibody specific for TSH. Standards or samples are then added to the microtiter plate wells and TSH if present, will bind to the antibody pre-coated wells. In order to quantitatively determine the amount of TSH present in the sample, a standardized preparation of horseradish peroxidase (HRP)-conjugated polyclonal antibody, specific for TSH are added to each well to �sandwich� the TSH immobilized on the plate. The microtiter plate undergoes incubation, and then the wells are thoroughly washed to remove all unbound components. Next, a TMB (3,3',5,5' tetramethyl-benzidine) substrate solution is added to each well. The enzyme (HRP) and substrate are allowed to react over a short incubation period. Only those wells that contain TSH and enzyme-conjugated antibody will exhibit a change in colour. The enzyme-substrate reaction is terminated by the addition of a sulfuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm. In order to measure the concentration of TSH in the sample, this Human TSH ELISA Kit includes a set of calibration standards (6 standards). The calibration standards are assayed at the same time as the samples and allow the operator to produce a standard curve of Optical Density (O.D.) versus TSH concentration (uIU/ml). The concentration of TSH in the samples is then determined by comparing the O.D. of the samples to the standard curve. Sensitivity: <0.2uIU/ml Range: 0.2-40uIU/ml Kit Components: T5400-01B1: Microtiter Plate, 1x 96 wells T5400-01B12: Conjugate, 1x 12ml T5400-01B13. Standard: 40uIU/ml, 1x1vial T5400-01B14. Standard: 20uIU/ml, 1x1vial T5400-01B15. Standard: 10uIU/ml, 1x1vial T5400-01B16. Standard: 5uIU/ml., 1x1vial T5400-01B17. Standard: 1uIU/ml, 1x1vial T5400-01B18. Standard: 0uIU/ml, 1x1vial T5400-01B19. Substrate A, 1x10ml T5400-01B110. Substrate B, 1x10ml T5400-01B111. Stop Solution, 1x7ml. 2N sulfuric acid. Materials required but not provided: 1. Single or multi-channel precision pipettes with disposable tips: 10-10ul and 50-200ul for running the assay. 2. Pipettes: 1ml, 5ml, and 10ml for reagent preparation. 3. Multi-channel pipette reservoir or equivalent reagent container. 4. Test tubes and racks. 5. Polypropylene tubes or containers (25ml). 6. Incubator (37�2 degrees C) 7. Microtiter plate reader (450 nm�2 nm) 8. Automatic microtiter plate washer or squirt bottle 9. Sodium hypochlorite solution, 5.25% (household liquid bleach). 10. Deionized or distilled water 11. Plastic plate cover 12. Disposable gloves 13. Absorbent paper Storage and Stability: Store components at 4 degrees C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.�