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Human IFN-β(Interferon Beta) ELISA Kit
Item No:
E-EL-H0085c
Chinese Name:
Alias:
GDF-15, MIC-1, MIC1, NAG-1, PDF, PLAB, PTGFB, TGF-PL
Keywords:
Classification:
Retail Price
¥ 1650
Market Price
¥ 1650
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Spec
- 48T
- 96T
- 96T*5
Inventory surplus
2997
隐藏域元素占位
- Product Comparison
Describe
Use
This kit is used for the in vitro quantification of IFN-β in human serum, plasma or other biological fluids.
Principle of detection
This kit is based on the double antibody sandwich ELISA method. Anti-Human IFN-β antibody is used to coat the enzyme plate, and during the experiment, Human IFN-β in the sample or standard will bind to the coating antibody, and the free component will be washed away. Biotinylated anti-human IFN-β antibody and horseradish peroxidase-labeled affin were added sequentially. The anti-human IFN-β antibody binds to the human IFN-β bound to the encapsulated antibody, biotin binds specifically to the affin and forms an immune complex, and the free component is washed away. A chromogenic substrate (TMB) is added, and TMB appears blue catalyzed by horseradish peroxidase and turns yellow after the addition of termination solution. The concentration of IFN-β in the sample was calculated by plotting the standard curve.
| Reaction Type | Sandwich method |
| Specification | 96T |
| Reaction time | 3.5h |
| Reactivity | Human |
| Detection method | Colormetric |
| Detection range | 31.25—2000 pg/mL |
| Sensitivity | 18.75 pg/mL |
| Sample volume | 100μL |
| Sample type | Serum, plasma or other biological fluids |
Specificity
Detects human IFN-β in samples with no significant cross-reactivity with other related proteins.
Repeatability
Intra-plate and inter-plate coefficients of variation are <10%.
Typical data
The OD values of the standard curves may vary due to different operating conditions (e.g. operator, pipetting technique, plate washing technique and stabilization conditions). The following data and curves are for reference only. Experimenters need to establish standard curves according to their own experiments.
| (pg/mL) | O.D | Average | Corrected |
|---|---|---|---|
| 2000 | 2.51 2.522 |
2.516 | 2.439 |
| 1000 | 1.671 1.725 |
1.698 | 1.621 |
| 500 | 0.978 0.948 |
0.963 | 0.886 |
| 250 | 0.461 0.473 |
0.467 | 0.39 |
| 125 | 0.281 0.263 |
0.272 | 0.195 |
| 62.5 | 0.183 0.173 |
0.178 | 0.101 |
| 31.25 | 0.12 0.138 |
0.129 | 0.052 |
| 0 | 0.070 0.084 |
0.077 | -- |
Precision
Intra-plate precision: 20 times on 1 plate for low, medium and high concentration samples.
Intra-plate precision: 20 times on 3 plates for low, medium and high concentrations.
| Intra-assay Precision | Inter-assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) |
105.20 | 289.60 | 951.90 | 107.40 | 261.00 | 1001.00 |
| Standard deviation |
5.90 | 14.20 | 40.00 | 5.40 | 11.70 | 54.10 |
| C V (%) | 5.61 | 4.90 | 4.20 | 5.03 | 4.48 | 5.40 |
Recovery
Recovery experiments were performed by adding known concentrations of target proteins to five different samples, and the recovery range and average recovery were obtained.
| Sample Type | Range (%) | Average Recovery (%) |
|---|---|---|
| Serum (n=8) | 88-102 | 93 |
| EDTA plasma (n=8) | 92-106 | 98 |
| Cell culture media (n=8) | 86-98 | 93 |
Linearity
Recovery experiments were performed by adding known concentrations of target proteins to 5 samples, and the range of recovery rates and average recovery rates were obtained. Five samples were diluted 2, 4, 8, and 16 times for recovery experiments, and the range of recovery rates and average recovery rates were obtained.
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
|---|---|---|---|---|
| 1:2 | Range (%) | 94-107 | 91-106 | 88-99 |
| Average (%) | 101 | 97 | 94 | |
| 1:4 | Range (%) | 100-117 | 84-99 | 96-112 |
| Average (%) | 107 | 90 | 103 | |
| 1:8 | Range (%) | 98-113 | 80-96 | 95-110 |
| Average (%) | 104 | 87 | 102 | |
| 1:16 | Range (%) | 98-116 | 82-92 | 96-110 |
| Average (%) | 106 | 87 | 103 |
Kit composition and storage
Unopened kits can be stored at 2-8°C for one week; if the kit is used after one week, please unpack the kit and store each component separately according to the conditions in the table below.
| Chinese Name | English Name | Specification | Storage conditions |
|---|---|---|---|
| ELISA酶标板(可拆卸) | Micro ELISA Plate(Dismountable) | 8 holes×12 strips | -20℃, can be stored for 6 months |
| 冻干标准品 | Reference Standard | 2 sticks | |
| 浓缩生物素化抗体(100×) | Concentrated Biotinylated Detection Ab | 1 x 120 μL | |
| 浓缩HRP酶结合物(100×) | Concentrated HRP Conjugate | 1 x 120 μL | -20℃(keep away from light), can be stored for 6 months |
| 标准品&样品稀释液 | Reference Standard & Sample Diluent | 1 bottle of 20 mL | 2-8℃. Can be stored for 6 months |
| 生物素化抗体稀释液 | Biotinylated Detection Ab Diluent | 1 bottle 14 mL | |
| 酶结合物稀释液 | HRP Conjugate Diluent | 1 bottle 14 mL | |
| 浓缩洗涤液(25×) | Concentrated Wash Buffer (25×) | 1 bottle 30 mL | |
| 底物溶液(TMB) | Substrate Reagent | 1 bottle 10 mL | 2-8℃. Can be stored for 6 months |
| 反应终止液 | Stop Solution | 1 bottle 10 mL | 2-8℃ |
| 封板覆膜 | Plate Sealer | 5 sheets | |
| 产品说明书 | Product Description | 1 copy | |
| 质检报告 | Certificate of Analysis | 1 copy |
Note: All reagent bottles must be tightly capped to prevent evaporation and microbial contamination.
The volume of reagents is subject to the actual shipping instructions. The relevant reagents will be slightly more than the volume indicated on the label when dispensing, please measure rather than pour directly when using.
Items required for the test
1. enzyme calibrator (450nm wavelength filter)
2. High precision pipettes, EP tubes and disposable tips: 0.5-10 μL, 2-20 μL, 20-200 μL, 200-1000 μL
3. 37°C thermostat, double-distilled water or deionized water
4. absorbent paper
Operation steps
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1. Add 100 μL of standard working solution or sample to the corresponding plate wells and incubate at 37°C for 90 minutes |
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2. Immediately after discarding the liquid in the plate, add 100 μL of biotinylated antibody working solution and incubate at 37°C for 60 minutes |
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3. Discard the liquid in the plate and wash the plate 3 times |
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4. Add 100μL of HRP enzyme conjugate working solution to each well, incubate at 37℃ for 30 minutes, discard the liquid in the plate, and wash the plate 5 times |
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5. Add 90μL of substrate solution to each well and incubate at 37℃ for 15 minutes |
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6. Add 50μL of termination solution to each well |
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7. Read immediately at 450 nm and process the data |
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