fT4(Free Thyroxine) ELISA Kit (E-EL-0122)
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For research use only.
Product Summary
| Sensitivity | 0.94 pg/mL |
| Detection Range | 1.56-100 pg/mL |
| Sample Volume | 50 μL |
| Total Assay Time | 2 h 30 min |
| Reactivity | Universal |
| Specificity | This kit recognizes Universal fT4 in samples.No significant cross-reactivity or interference between Universal fT4 and analogues was observed |
| Recovery | 80%-120% |
| Sample Type | Human: Serum, EDTA plasma, heparinized plasma, Saliva, Breast milk, Urine and Stool; Mouse: Serum, EDTA plasma, Urine and Tissue homogenate; Rat: Serum, EDTA plasma, Urine and Tissue homogenate; Cell lysate, Cell culture supernatant. |
| Detection Method | Colorimetric method, ELISA, Competitive |
| Assay Type | Competitive-ELISA |
| Size | 96T / 48T / 24T / 96T*5 / 96T*10 |
| Storage | 2-8℃ |
| Expiration Date | 12 months |
Performance
| Typical data |
The following data was generated by the Quality Control Department, under controlled laboratory conditions (ambient temperature: 18-25 °C, relative humidity: 35-75%) using standardized procedures (TMB reaction at 37 °C in the dark for 15 minutes, followed by termination and OD measurement). These values are provided for reference only.
Actual results may vary due to differences in laboratory conditions, operator technique, and equipment. Users are required to generate a standard curve using their own experimental data.
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| Precision |
Intra-assay Precision (Within-run Precision): Three samples representing low, mid, and high concentrations of Universal fT4 were tested 20 times on a single plate.
Inter-assay Precision (Between-run Precision): Three samples representing low, mid, and high concentrations of Universal fT4 were tested on three separate plates, with 20 replicates per plate, to assess variability among assays.
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| Recovery |
The recovery of Universal fT4 was evaluated by spiking samples at low, mid, and high concentrations across the assay range in various sample matrices.
The assay performance was assessed by comparing the measured concentrations to the expected spiked amounts to determine the percent recovery.
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| Linearity |
Linearity of the assay was evaluated by spiking samples with high concentrations of Universal fT4 and performing serial dilutions using Standard & Sample Diluent to produce concentrations spanning the assay's dynamic range.
The measured values were then compared to the expected concentrations to assess the linearity of response.
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| Stability |
Each kit batch is subjected to accelerated stability testing and real-time stability monitoring. Sample performance is evaluated after storage at 37 °C for 10 days to assess the impact of elevated temperature on assay reliability and reagent integrity.
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Test Principle
This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with Universal fT4. During the reaction, Universal fT4 in the sample or standard competes with a fixed amount of Universal fT4 on the solid phase supporter for sites on the Biotinylated Detection Ab specific to Universal fT4. Excess conjugate and unbound sample or standard are washed away, and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns from blue to yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of Universal fT4 in tested samples can be calculated by comparing the OD of the samples to the standard curve.
Background
Free thyroxine (fT4) is the fraction of serum thyroxine that is not bound to carrier proteins and is therefore directly available to tissues. It accounts for only a small proportion of total circulating thyroxine, and its measurement is a core component of routine thyroid testing. Because it is not confounded by changes in binding protein concentration, it reflects functional thyroid status more accurately than total T4.
Free T4 represents the immediately available pool of thyroid secretory output, maintained within a narrow range by the negative feedback of the hypothalamus-pituitary-thyroid axis. Measured together with thyroid-stimulating hormone, it distinguishes overt from subclinical hyperthyroidism and hypothyroidism and avoids the misinterpretation caused by altered binding proteins in pregnancy, estrogen therapy or non-thyroidal illness.
Measurement of fT4 is used to diagnose and monitor thyroid disease. In hypothyroidism it generally falls before free T3 and is therefore the preferred marker, and it is also used to monitor suppressive therapy and to assess thyroid status during pregnancy and critical illness, providing a binding-protein-independent index of tissue-available hormone in clinical and research settings.
| Research Area | DevelopmentalBiology , SignalTransduction |
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