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Human IL-3(Interleukin 3) ELISA Kit
Item No:
E-EL-H0100c
Chinese Name:
Alias:
IL3, MCGF, MCSF, MULTI-CSF, P-Cell Stimulating Factor
Keywords:
Classification:
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隐藏域元素占位
- Product Comparison
Describe
Application
This kit is used for the in vitro quantification of IL-3 in human serum, plasma or other biological fluids
Principle of detection
This kit adopts double antibody sandwich ELISA method. Anti-Human IL-3 antibody is used to coat the ELISA plate. Human IL-3 in the sample or standard will bind to the coating antibody during the experiment, and the free components are washed away. Biotinylated anti-human IL-3 antibody and horseradish peroxidase-labeled affinity factor are added sequentially. The anti-human IL-3 antibody binds to the human IL-3 bound to the encapsulated antibody, and the biotin binds specifically to the affin to form an immune complex, and the free components are 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 IL-3 in the sample was calculated by plotting the standard curve, using an enzyme marker to measure the OD value at 450 nm, with a positive ratio between IL-3 concentration and OD450 value.
| Reaction Type | Sandwich method |
| Specification | 96T |
| Reaction time | 3.5h |
| Reactivity | Human |
| Detection method | Colormetric |
| Detection range | 15.63—1000 pg/mL |
| Sensitivity | 9.38 pg/mL |
| Sample volume | 100μL |
| Sample type | Serum, plasma or other biological fluids |
Specificity
Detects human IL-3 in samples with no significant cross-reactivity with other related proteins.
Repeatability
Intra-plate and inter-plate coefficients of variation <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 |
|---|---|---|---|
| 1000 | 2.418 2.426 |
2.422 | 2.342 |
| 500 | 1.647 1.655 |
1.651 | 1.571 |
| 250 | 0.993 0.967 |
0.98 | 0.9 |
| 125 | 0.46 0.464 |
0.462 | 0.382 |
| 62.5 | 0.263 0.259 |
0.261 | 0.181 |
| 31.25 | 0.181 0.181 |
0.181 | 0.101 |
| 15.63 | 0.127 0.137 |
0.132 | 0.052 |
| 0 | 0.077 0.083 |
0.08 | -- |
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) |
47.04 | 159.61 | 356.96 | 47.69 | 156.29 | 343.47 |
| Standard deviation |
2.63 | 8.16 | 13.24 | 3.31 | 7.64 | 13.19 |
| C V (%) | 5.59 | 5.11 | 3.71 | 6.94 | 4.89 | 3.84 |
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) | 86-96 | 91 |
| EDTA plasma (n=8) | 88-104 | 95 |
| Cell culture media (n=8) | 86-100 | 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 (%) | 93-111 | 92-102 | 94-108 |
| Average (%) | 101 | 97 | 100 | |
| 1:4 | Range (%) | 89-102 | 84-96 | 87-101 |
| Average (%) | 94 | 88 | 92 | |
| 1:8 | Range (%) | 86-100 | 83-95 | 86-96 |
| Average (%) | 93 | 90 | 91 | |
| 1:16 | Range (%) | 88-99 | 79-93 | 87-101 |
| Average (%) | 93 | 86 | 94 |
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 | Preservation conditions |
|---|---|---|---|
| ELISA酶标板(可拆卸) | Micro ELISA Plate(Dismountable) | 8 holes x 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℃(avoid light), can be stored for 6 months |
| 标准品&样品稀释液 | Reference Standard & Sample Diluent | 1 bottle 20 mL | 2-8°C. 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℃(avoid light) |
| 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 sticks | |
| 质检报告 | Certificate of Analysis | 1 sticks |
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.
Self-provided items required for the test
1. ELISA (450 nm 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 100 μL of HRP enzyme conjugate working solution to each well, incubate for 30 minutes at 37°C, discard the liquid in the plate, and wash the plate 5 times. |
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5. Add 90 μL of substrate solution to each well and incubate at 37°C for about 15 minutes |
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6. Add 50 μL of termination solution per well |
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7. Read immediately at 450nm wavelength and process the data |
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