ELISA Kit for Tetrahydrobiopterin (THB)
BH4; Kuvan; Sapropterin
- Product No.CEG421Ge
- Organism SpeciesPan-species (General) Same name, Different species.
- Test MethodCompetitive Inhibition
- Assay Length2h
- Detection Range391-100,000pg/mL
- SensitivityThe minimum detectable dose of this kit is typically less than 137pg/mL.
- Sample Typeserum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
- Download Instruction Manual
- UOM 48T96T 96T*5 96T*10 96T*100
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FOB
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Packages (Simulation)
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Packages (Simulation)
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Results demonstration
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Typical Standard Curve
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ISO9001: 2008, ISO13485: 2003 Registered
Specificity of the ELISA Kit for Tetrahydrobiopterin (THB)
This assay has high sensitivity and excellent specificity for detection of Tetrahydrobiopterin (THB).
No significant cross-reactivity or interference between Tetrahydrobiopterin (THB) and analogues was observed.
Recovery of the ELISA Kit for Tetrahydrobiopterin (THB)
Matrices listed below were spiked with certain level of Tetrahydrobiopterin (THB) and the recovery rates were calculated by comparing the measured value to the expected amount of Tetrahydrobiopterin (THB) in samples.
| Matrix | Recovery range (%) | Average(%) |
| serum(n=5) | 89-104 | 93 |
| EDTA plasma(n=5) | 89-98 | 94 |
| heparin plasma(n=5) | 82-94 | 88 |
Precision of the ELISA Kit for Tetrahydrobiopterin (THB)
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Tetrahydrobiopterin (THB) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Tetrahydrobiopterin (THB) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%
Linearity of the ELISA Kit for Tetrahydrobiopterin (THB)
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Tetrahydrobiopterin (THB) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
| Sample | 1:2 | 1:4 | 1:8 | 1:16 |
| serum(n=5) | 96-105% | 86-101% | 95-104% | 79-95% |
| EDTA plasma(n=5) | 95-104% | 78-95% | 79-98% | 78-96% |
| heparin plasma(n=5) | 93-105% | 80-91% | 93-104% | 86-101% |
Stability of the ELISA Kit for Tetrahydrobiopterin (THB)
The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.
Assay procedure summary of the ELISA Kit for Tetrahydrobiopterin (THB)
1. Prepare all reagents, samples and standards;
2. Add 50µL standard or sample to each well.
And then add 50µL prepared Detection Reagent A immediately.
Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.
Test principle of the ELISA Kit for Tetrahydrobiopterin (THB)
This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to Tetrahydrobiopterin (THB) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled Tetrahydrobiopterin (THB) and unlabeled Tetrahydrobiopterin (THB) (Standards or samples) with the pre-coated antibody specific to Tetrahydrobiopterin (THB). After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of Tetrahydrobiopterin (THB) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of Tetrahydrobiopterin (THB) in the sample.
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