6-Sulfatoxymelatonin ELISA

Product catalog summary

Intended Use
The ALPCO 6-Sulfatoxymelatonin ELISA is designed for the quantitative measurement of 6-sulfatoxymelatonin in human urine samples. It is strictly for research purposes and not for diagnostic use.

Principle of the Assay
This ELISA is based on a competitive binding principle where an unknown amount of antigen in the sample competes with a fixed amount of enzyme-labeled antigen for antibody binding sites. The color intensity developed is inversely proportional to the antigen concentration.

Materials Supplied
The kit includes a 96-well microplate, calibrators, controls, wash buffer concentrate, melatonin sulfate antiserum, enzyme conjugate, assay buffer, TMB substrate solution, and stop solution. All components are ready to use except the wash buffer and enzyme conjugate, which require dilution.

Materials Required
Additional materials needed include precision pipettes, a microplate washer, a microplate reader, a centrifuge, and a vortex.

Precautions
The kit is for research use only. Users should follow safety guidelines, avoid mixing reagents from different lots, and handle all chemicals as hazardous waste. Proper lab attire is recommended.

Storage Conditions
The kit should be stored at 2-8°C, away from heat and direct sunlight. The microtiter strips are stable until the kit's expiry date if stored properly.

Sample Handling
Urine samples can be spontaneous or 24-hour collections. They should be mixed and centrifuged before use. Samples can be stored at 2-8°C for up to 4 days or at ≤-20°C for up to 15 years.

Reagent Preparation
The wash buffer concentrate must be diluted 1:20 with distilled water, and the enzyme conjugate must be diluted 1:41 with assay buffer immediately before use.

Quality Control
Control samples should be analyzed with each run, and results should be evaluated using statistical methods to ensure validity.

Assay Procedure
The procedure involves pipetting calibrators, controls, and samples into wells, adding assay buffer, enzyme conjugate, and antiserum, followed by incubation, washing, and substrate addition. Optical density is measured at 450 nm.

Calculation of Results
Results are calculated using a calibrator curve plotted on semi-logarithmic graph paper or using automated methods. The concentration is read directly from the curve.

Performance Characteristics
The assay has a sensitivity of 1.0 ng/mL. Precision is indicated by intra-assay and inter-assay CVs ranging from 5.2% to 14.9%.

Specificity
The assay shows 100% cross-reactivity with melatonin sulfate and negligible cross-reactivity with other substances.

Catalog excerpts

6-Sulfatoxymelatonin ELISA Immunoassay for the quantitative determination of 6sulfatoxymelatonin in human urine. For Research Use Only. Not For Use In Diagnostic Procedures. Catalog Number: Size: Version:

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The ALPCO 6-Sulfatoxymelatonin ELISA is intended for the quantitative determination of 6-sulfatoxymelatonin in human urine samples. For Research Use Only. Not for use in diagnostic procedures. PRINCIPLE OF THE ASSAY Solid phase enzyme-linked immunosorbent assay (ELISA) based on the competition principle. An unknown amount of antigen present in the sample and a fixed amount of enzyme labelled antigen compete for the binding sites of the antibodies coated onto the wells. After incubation the wells are washed to stop the competition reaction. After the substrate reaction the intensity of the developed...

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Precision pipettes for dispensing up to 1000 μL (with disposable tips) Repeating or multi-channel pipette for dispensing up to 1000 μL Volumetric containers and pipettes for reagent preparation Distilled/Deionized water for reagent preparation Microplate washer or wash bottle Microplate shaker capable of 700-900 rpm Microplate reader Centrifuge (1,000 x g to 3,000 x g) Vortex for sample preparation PRECAUTIONS 1. For research use only. Not for use in diagnostic procedures. 2. Before starting the assay, read the instructions completely and carefully. Use the valid version of the package insert...

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17. Microtiter plate washing is important. Improperly washed wells will give erroneous results. It is recommended to use a multichannel pipette or an automatic microtiter plate washing system. Do not allow the wells to dry between incubations. Do not scratch coated wells during rinsing and aspiration. Rinse and fill all reagents with care. While rinsing, check that all wells are filled precisely with Wash Buffer, and that there are no residues in the wells. 18. Humidity affects the coated wells/tubes. Do not open the pouch until it reaches room temperature. Unused wells/tubes should be returned...

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QUALITY CONTROL Control samples should be analyzed with each run. Results generated from the analysis of control samples, should be evaluated for acceptability using appropriate statistical methods. The results for the samples may not be valid if within the same assay one or more values of the quality control sample are outside the acceptable limits. ASSAY PROCEDURE Gently mix all reagents before use. 1. Pipette 10 µL of each Calibrator, Control and urine sample into polystyrene, polypropylene or glass tubes. Avoid direct sun light. 2. Pipette 500 µL of Assay Buffer into each tube. Vortex. 3....

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PERFORMANCE CHARACTERISTICS Analytical Sensitivity LOD: 1.0 ng/mL Typical Calibration Curve (Example. Do not use for calculation) Melatonin Sulfate (ng/mL) Range (ng/mL) Dilutional Linearity Range (ng/mL) Serial dilution up to Range (%) 1:32 Spiking Recovery Mean recovery after spiking (%) Range (%) 105.8

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Cross-reactivity of other substances tested < 0.0001 %

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