**Porcine Interleukin-17 (IL-17) ELISA Kit – For Research Use Only**
This ELISA kit is designed for the quantitative in vitro determination of IL-17 levels in porcine serum, plasma, and other biological fluids. The kit provides a reliable and accurate method to measure IL-17 concentrations using a sandwich ELISA technique. The expiration date is 6 months from the date of receipt, and it should be stored at 2–8°C.
**Principle of the Assay**
The kit employs a double-antibody sandwich method. A microtiter plate is coated with purified porcine IL-17 antibody to form a solid-phase antibody. After incubation with the sample, IL-17 binds to the immobilized antibody. An HRP-conjugated secondary antibody is then added, forming an immune complex. TMB substrate is used for color development, which turns blue under the action of HRP and changes to yellow upon addition of the stop solution. The optical density (OD) is measured at 450 nm, and the IL-17 concentration is determined by comparing the OD values with a standard curve.
**Kit Components**
- 96-well microtiter plate
- 1×96 strip plate (coated with anti-IL-17 antibody)
- 1 user manual
- 2 sealing films
- 1 sealed bag
- 1 standard (270 pg/ml, 0.5 ml)
- 1 standard diluent (1.5 ml)
- 1 HRP-conjugate reagent (6 ml)
- 1 sample diluent (6 ml)
- 1 Chromogen Solution A (6 ml)
- 1 Chromogen Solution B (6 ml)
- 1 Stop Solution (6 ml)
- 1 concentrated washing solution (20 ml × 30 times)
**Sample Preparation**
- **Serum**: Allow blood to clot at room temperature for 10–20 minutes, centrifuge at 2000–3000 rpm for 20 minutes, and collect the supernatant.
- **Plasma**: Use EDTA or citrate as anticoagulant. Mix for 10–20 minutes, centrifuge, and collect supernatant.
- **Urine**: Collect in a sterile container, centrifuge, and remove supernatant.
- **Cell culture supernatant**: Centrifuge, discard cells, and use the supernatant.
- **Tissue samples**: Homogenize in PBS, centrifuge, and collect the supernatant.
- **Note**: Avoid samples containing NaN3, as it inhibits HRP activity.
**Assay Procedure**
1. Prepare a serial dilution of the standard.
2. Add samples and standards to the wells.
3. Incubate the plate at 37°C for 30 minutes.
4. Wash the plate 5 times with diluted wash buffer.
5. Add HRP-conjugated reagent to each well except blank.
6. Incubate again for 30 minutes.
7. Wash the plate again.
8. Add TMB substrates and incubate for 15 minutes in the dark.
9. Stop the reaction by adding stop solution.
10. Measure absorbance at 450 nm within 15 minutes.
**Important Notes**
- Allow the kit to equilibrate at room temperature before use.
- Store unused strips in a sealed bag.
- Avoid light exposure during the color development step.
- Always include blank wells and perform duplicate measurements.
- If the sample OD exceeds the highest standard, dilute the sample and recalculate.
- Do not mix reagents from different batches.
**Performance**
- Correlation coefficient (R) of the standard curve: ≥0.92
- Intra-assay variation: <9%
- Inter-assay variation: <15%
- Detection range: 3.8 pg/ml – 200 pg/ml
**Company Information**
Zhengzhou Weier Biotechnology Co., Ltd. is a comprehensive enterprise specializing in research, production, and sales of biological products. Our product line includes ELISA kits, antibodies, standards, cell culture media, and more. We offer high-quality solutions for researchers worldwide. Welcome to inquire about our products!
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