Melatonin Sulfate (MS) ELISA Kit

Antigen: Melatonin Sulfate (MS)

Reaktivität: Human

Applikation: ELISA

Zertifikate: ISO 9001:2008

Produktnummer: ABIN366939

Produktbeschreibung

Produktmerkmale: This immunoassay kit allows for the in vitro quantitative determination of human MS concentrations in serum, plasma and other biological fluids.

Proben: Serum, Plasma

Spezifität: This assay recognizes MS.

Anwendungen

Prinzip: This assay employs the direct competitive inhibition enzyme immunoassay technique. An antibody specific for MS has been pre-coated onto a microplate. Add MS (Standards or samples) to the well, and then add Avidin conjugated MS. A competitive inhibition reaction is launched between MS (Standards or samples) and Avidin conjugated MS with the MS antibody. Then add the HRP-avidin to each well. The substrate solutions are added to the wells, respectively. And the color develops in opposite to the amount of MS bound in the initial step. The color development is stopped and the intensity of the color is measured.

Protokoll: Preparation of Reagents: Bring all reagents to room temperature before use. 1. Wash Buffer If crystals have formed in the concentrate, warm up to room temperature and mix gently until the crystals have completely dissolved. Dilute 20 ml of Wash Buffer Concentrate into deionized or distilled water to prepare 500 ml of Wash Buffer. 2. Standard Reconstitute the Standard with 1.0 ml of Sample Diluent. This reconstitution produces a stock solution of 200 ng/ml. Allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making serial dilutions. The undiluted standard serves as the high standard (200 ng/ml). The Sample Diluent serves as the zero standard (0 pg/ml). 3. Biotin-MS Dilute to the working concentration using Biotin-MS Diluent(1:100), respectively. 4. HRP-avidin Dilute to the working concentration using HRP-avidin Diluent(1:100), respectively. Precaution: The Stop Solution provided with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material. Sample Collection and Storage: Assay Procedure: Calculation of Results: Average the duplicate readings for each standard, control, and sample and divide the average zero standard optical density. Create a standard curve by reducing the data using computer software. As an alternative, construct a standard curve by plotting the absorbance ratio for each standard on the y-axis against the concentration on the x-axis and draw a best fit curve through the points on the graph. The data may be linearized by plotting the log of the MS concentrations versus the ratio and the best fit line can be determined by regression analysis. This procedure will produce an adequate but less precise fit of the data. If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor. Limitations of the Procedure: The kit should not be used beyond the expiration date on the kit label. Do not mix or substitute reagents with those from other lots or sources. It is important that the Calibrator Diluent selected for the standard curve be consistent with the samples being assayed. If samples generate values higher than the highest standard, dilute the samples with the appropriate diluent and repeat the assay. Any variation in operator, pipetting technique, washing technique, incubation time or temperature, and kit age can cause variation in binding. This assay is designed to eliminate interference by soluble receptors, binding proteins, and other factors present in biological samples. Until all factors have been tested in the Immunoassay, the possibility of interference cannot be excluded.

Testdurchführung: Assay Time: 1-3h
Sample Volume: 50-100ul

Applikationshinweise: Detection Wavelength: 450 nm

Bestandteile: Reagent Quantity Assay plate 1 Standard 2 Sample Diluent 1 x 20 ml Biotin-MS Diluent 1 x 10 ml HRP-avidin Diluent 1 x 10 ml Biotin-MS 1 x 120µl HRP-avidin 1 x 120µl Wash Buffer 1 x 20 ml (25×concentrate) TMB Substrate 1 x 10 ml Stop Solution 1 x 10 ml

Lagerung: 1. Unopened test kits should be stored at 2-8°C upon receipt and the microtiter plate should be kept in a sealed bag. The test kit may be used throughout the expiration date of the kit. Refer to the package label for the expiration date. 2. Opened test kits will remain stable until the expiring date shown, provided it is stored as prescribed above. 3. A microtiter plate reader with a bandwidth of 10 nm or less and an optical density range of 0-3 OD or greater at 450nm wavelength is acceptable for use in absorbance measurement. Expiration date: six months from the date of manufacture.

Beschränkungen: Nur für Forschungszwecke einsetzbar