Rat IL-4(Interleukin 4) ELISA Kit
Item No:
E-EL-R0014c
Chinese Name:
Alias:
IL4, BCGF-1, BCGF1, BSF-1, BSF1
Keywords:
Classification:
Retail Price
¥ 2400
Market Price
¥ 2400
-
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 IL-4 in Rat serum, plasma or other biological fluids.
Detection principle
This kit adopts double antibody sandwich ELISA method. Anti-Rat IL-4 antibody is used to coat the ELISA plate. The rat IL-4 in the sample or standard will bind to the coating antibody during the experiment, and the free components are washed away. Biotinylated anti-rat IL-4 antibody and horseradish peroxidase-labeled affin were added sequentially. The anti-rat IL-4 antibody binds to the rat IL-4 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. The chromogenic substrate (TMB) was added, and the TMB appeared blue catalyzed by horseradish peroxidase and turned yellow after the addition of termination solution. The concentration of IL-4 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-4 concentration and OD450 value.
| Reaction Type | Sandwich method |
| Specification | 96T |
| Reaction time | 3.5h |
| Reactivity | Rat |
| 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 rat IL-4 in samples with no significant cross-reactivity with other related proteins.
Repeatability
Within the plates, the coefficients of variation between the plates were <10%.
Typical Data
The OD values of the standard curves may vary due to different experimental operating conditions (e.g. operator, pipetting technique, plate washing technique and stabilization conditions, etc.). 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 | 2.386 2.402 |
2.394 | 2.341 |
| 500 | 1.642 1.682 |
1.662 | 1.609 |
| 250 | 0.946 0.922 |
0.934 | 0.881 |
| 125 | 0.402 0.43 |
0.416 | 0.363 |
| 62.5 | 0.245 0.233 |
0.239 | 0.186 |
| 31.25 | 0.158 0.152 |
0.155 | 0.102 |
| 15.63 | 0.103 0.107 |
0.105 | 0.052 |
| 0 | 0.053 0.053 |
0.053 | -- |

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) |
47.20 | 134.60 | 394.80 | 47.80 | 134.90 | 433.70 |
| Standard deviation |
3.20 | 6.50 | 15.40 | 2.50 | 5.90 | 21.30 |
| C V (%) | 6.78 | 4.83 | 3.90 | 5.23 | 4.37 | 4.91 |
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) | 91-102 | 97 |
| EDTA plasma (n=8) | 86-99 | 93 |
| Cell culture media (n=8) | 91-103 | 96 |
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 (%) | 88-103 | 94-110 | 86-101 |
| Average (%) | 95 | 102 | 92 | |
| 1:4 | Range (%) | 102-117 | 81-93 | 98-112 |
| Average (%) | 107 | 87 | 105 | |
| 1:8 | Range (%) | 98-113 | 84-95 | 95-109 |
| Average (%) | 106 | 88 | 101 | |
| 1:16 | Range (%) | 96-110 | 84-96 | 95-110 |
| Average (%) | 104 | 90 | 101 |
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 enzyme standard plate (detachable) | Micro ELISA Plate(Dismountable) | 8 holes×12 strips | -20℃, can be stored for 6 months |
| Lyophilized standards | Reference Standard | 2 sticks | |
| Concentrated biotinylated antibody(100×) | Concentrated Biotinylated Detection Ab | 1 x 120 μL | |
| Concentrated HRP enzyme conjugate(100×) | Concentrated HRP Conjugate | 1 x 120 μL | -20℃(keep away from light), can be stored for 6 months |
| Standard & Sample Diluent | Reference Standard & Sample Diluent | 1 bottle of 20 mL | 2-8℃. Can be stored for 6 months |
| Biotinylated antibody diluent | Biotinylated Detection Ab Diluent | 1 bottle 14 mL | |
| Enzyme Conjugate Diluent | HRP Conjugate Diluent | 1 bottle 14 mL | |
| Concentrated Washing Solution(25×) | Concentrated Wash Buffer (25×) | 1 bottle 30 mL | |
| Substrate solution (TMB) | Substrate Reagent | 1 bottle 10 mL | 2-8℃(keep away from light) |
| Reaction termination solution | Stop Solution | 1 bottle 10 mL | 2-8℃ |
| Plate lamination | Plate Sealer | 5 sheets | |
| Product manual | Product Description | 1 copy | |
| Quality inspection report | 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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