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Human TNF-α (Tumor Necrosis Factor Alpha) CLIA Kit
SKU: E-CL-H0108
Human TNF-α (Tumor Necrosis Factor Alpha) CLIA Kit
| Detection Range | 7.81-500 pg/mL |
| Sensitivity | 4.69 pg/mL |
Product Details
Properties
| Assay type | Sandwich-CLIA |
| Format | 96T |
| Assay time | 3.5h |
| Detection range | 7.81-500 pg/mL |
| Sensitivity | 4.69 pg/mL |
| Sample type &Sample volume | serum, plasma and other biological fluids; 100μL |
| Specificity | This kit recognizes Human TNF-α in samples. No significant cross-reactivity or interference between Human TNF-α and analogues was observed. |
| Reproducibility | Both intra-CV and inter-CV are < 15%. |
| Application | This CLIA kit applies to the in vitro quantitative determination of Human TNF-α concentrations in serum, plasma and other biological fluids. |
Test Principle
This CLIA kit uses the Sandwich-CLIA principle. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Human TNF-α. Standards or samples are added to the micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human TNF-α and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human TNF-α, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit (RLU) value is measured by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Human TNF-α. You can calculate the concentration of Human TNF-α in the samples by comparing the RLU value of the samples to the standard curve.
Kit Components and Storage
An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.
| Item | Specifications | Storage |
|---|---|---|
| Micro CLIA Plate(Dismountable) | 96T: 8 wells ×12 strips 48T: 8 wells ×6 strips |
-20℃, 6 months |
| Reference Standard | 96T: 2 vials 48T: 1 vial |
|
| Concentrated Biotinylated Detection Ab (100×) | 96T: 1 vial, 120 μL 48T: 1 vial, 60 μL |
|
| Concentrated HRP Conjugate (100×) | 96T: 1 vial, 120 μL 48T: 1 vial, 60 μL |
-20℃(Protect from light), 6 months |
| Reference Standard & Sample Diluent | 1 vial, 20 mL | 2-8°C, 6 months |
| Biotinylated Detection Ab Diluent | 1 vial, 14 mL | |
| HRP Conjugate Diluent | 1 vial, 14 mL | |
| Concentrated Wash Buffer (25×) | 1 vial, 30 mL | |
| Substrate Reagent A | 1 vial, 5 mL | 2-8℃(Protect from light) |
| Substrate Reagent B | 1 vial, 5 mL | |
| Plate Sealer | 5 pieces | |
| Manual | 1 copy | |
| Certificate of Analysis | 1 copy |
Technical Data:
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human TNF-α were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human TNF-α were tested on 3 different plates, 20 replicates in each plate.
| Intra-assay Precision | Inter-assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 25.60 | 58.03 | 169.01 | 27.58 | 54.45 | 158.50 |
| Standard deviation | 2.69 | 5.50 | 13.54 | 2.46 | 6.40 | 14.99 |
| CV (%) | 10.51 | 9.48 | 8.01 | 8.92 | 11.75 | 9.46 |
Recovery
The recovery of Human TNF-α spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
| Sample Type | Range (%) | Average Recovery (%) |
|---|---|---|
| Serum(n=8) | 100-113 | 107 |
| EDTA plasma(n=8) | 92-103 | 98 |
| Cell culture media(n=8) | 87-101 | 92 |
Target Information
| Synonyms | DIF, TNF-alpha, TNFA, TNFSF2 |
| Research Area | Cancer, Cardiovascular, Metabolism, Immunology, Signal transduction |
Assay Procedures
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1.Add 100 μL of standard or sample to each well. Incubate for 90 min at 37℃. |
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2.Remove the liquid. |
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3.Add 100 μL Biotinylated Detection Ab. Incubate 1 hour at 37℃. Aspirate and wash 3 times. |
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4.Add 100 μL HRP Conjugate. Incubate 30 min at 37℃. Aspirate and wash 5 times. |
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5.Add 100 μL Substrate Mixture Solution. Incubate for 5 min at 37℃. |
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6.Fluorescence appeared. Measure the RLU value with the Chemiluminescence immunoassay analyzer. Calculation of the results. |