Abraxis: 580002: 17-B Estradiol by Immunoassay, Magnetic Particle
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Official Method Name
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Abraxis 17-B Estradiol Assay Kit (100T) PN 580002 |
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Current Revision
| 2005 |
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Media
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WATER |
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Instrumentation
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Immunoassay |
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Method Subcategory
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Organic |
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Method Source
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Citation
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Brief Method Summary
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17B-Estradiol is detected using a colorimetric immunoassay (ELISA) procedure. A sample (0.25 mL) and paramagnetic particles attached with 17B-Estradiol-specific antibodies are added to a disposable test tube and allowed to incubate followed by the addition of an enzyme conjugate (enzyme-labeled 17B-Estradiol). Both the 17B-Estradiol in the sample and the enzyme conjugate compete for antibody binding sites on the paramagnetic particles in proportion to their concentrations. At the end of an incubation period, the magnetic particles are separated and washed with the aid of a magnetic separator. A substrate is then added which is catalyzed by the enzyme and converted from a colorless to a blue solution. The reaction is terminated with the addition of a dilute acid. The concentration of 17B-Estradiol in the sample is determined by measuring its absorbance at a specific wavelength (450 nm) using a photometer and comparing its absorbance to the absorbance of the calibrators. |
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Scope and Application
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This method determines 17B-Estradiol in water (groundwater, surface water, well water). |
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Applicable Concentration Range
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2.5 - 25.0 |
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Interferences
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Quality Control Requirements
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(A) Calibration with 3 standards and 1 blank, all analyzed in duplicate. (B) Precision: 3 matrix samples with different levels in the range for quantitative analysis analyzed daily for 5 days with 5 replicates in each of 5 assays. (C) Accuracy: 5 matrix samples spiked with the target analyte at 4 different levels in the range for quantitative analysis. (D) Validation: Analysis of 4 positive and 4 negative samples by an independent method for confirmation. |
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Sample Handling
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Samples are collected in glass containers with Teflon-lined caps; for compliance monitoring, samples should be held no longer than 14 days. If samples are held frozen, they can be held for longer periods of time. Immunoassay reagents are stored refrigerated until use. Samples containing gross particulate matter should be filtered using a 1 micron filter or centrifuged before use. Drinking water samples are typically dechlorinated with 0.008 percent sodium thiosulfate. |
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Maximum Holding Time
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14 days at 4 degrees C, longer if frozen |
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Relative Cost
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Less than $50 |
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Sample Preparation Methods
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