**Human Th17 ELISA Kit – For the quantitative in vitro determination of Human Th17 concentrations in serum, plasma, cerebrospinal fluid, tissue homogenate, and other body fluids. FOR LABORATORY RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES.**
Before using this kit, please read the entire package insert carefully. This ELISA (Enzyme-Linked Immunosorbent Assay) is designed for research purposes only and should not be used for diagnostic or clinical applications.
**INTENDED USE AND TEST PRINCIPLE**
The Human Th17 ELISA Kit is intended for laboratory research use only. The test is based on a sandwich immunoassay format. After incubation with the sample, a color change occurs due to the enzymatic reaction, which can be measured spectrophotometrically at 450 nm. A standard curve is generated by measuring the optical density (OD) of known concentrations of Th17, allowing the concentration of Th17 in unknown samples to be determined by comparison.
**SAMPLE COLLECTION AND STORAGE**
- **Serum**: Use a serum separator tube. Allow samples to clot for 2 hours at room temperature or overnight at 4°C before centrifugation at 2000×g for 20 minutes. Analyze immediately or store at -20°C. Avoid repeated freeze-thaw cycles.
- **Plasma**: Collect using heparin as an anticoagulant. Centrifuge within 30 minutes at 2000×g for 30 minutes at 2–8°C. Store at -20°C. Avoid freeze-thaw cycles.
- **Cell culture supernatants, tissue homogenates, and other biological fluids**: Remove particulates by centrifugation. Analyze immediately or store at -20°C. Ensure no hemolysis or granules are present in the sample.
**MATERIALS REQUIRED BUT NOT SUPPLIED**
1. Incubator set at 37°C
2. Microplate reader capable of measuring absorbance at 450 nm
3. Precision pipettes, disposable tips, and absorbent paper
4. Distilled or deionized water
**REAGENTS PROVIDED (Storage: 2–8°C)**
| Reagent | 96 Determinations | 48 Determinations |
|---------|-------------------|-------------------|
| Microplate (12×8 strips) | 1 | 1 |
| Standard (6 vials, 0.5 mL/vial) | 6 vials | 6 vials |
| Sample Diluent | 6.0 mL | 3.0 mL |
| HRP-Conjugate Reagent | 10.0 mL | 5.0 mL |
| 20X Wash Solution | 25 mL | 15 mL |
| Chromogen Solution A | 6.0 mL | 3.0 mL |
| Chromogen Solution B | 6.0 mL | 3.0 mL |
| Stop Solution | 6.0 mL | 3.0 mL |
| Closure Plate Membrane | 2 | 2 |
| User Manual | 1 | 1 |
| Sealed Bags | 1 | 1 |
*Note: Standard concentrations are 50, 25, 12.5, 6.25, 3.12, and 1.56 ng/mL. If sample values exceed the highest standard, dilute with Sample Diluent and repeat the assay.*
**PRECAUTIONS**
1. Do not mix reagents from different kit lots. All components are matched for optimal performance.
2. Allow all reagents to reach room temperature (20–25°C) before use. Avoid using water baths for thawing.
3. Do not use reagents beyond their expiration date.
4. Only use deionized or distilled water for reagent dilution.
5. Keep microplates in sealed bags until ready to use. Unused strips should be stored at 2–8°C with desiccant.
6. Use fresh pipette tips for each transfer to prevent contamination.
7. Do not use disposable knives in the assay, as no method guarantees complete safety from potential infectious agents.
8. Waste containing biological material must be treated with 1.0% sodium hypochlorite for at least 30 minutes before disposal.
9. Substrate solution may be contaminated; discard if it appears blue.
10. Substrate B contains 20% acetone—keep away from heat or flame.
11. Allow all reagents to warm to room temperature before use.
**REAGENT PREPARATION**
- **Wash Solution (1X)**: Dilute 1 volume of 20X Wash Solution with 19 volumes of deionized or distilled water. Store at 2–8°C for up to one month.
**ASSAY PROCEDURE**
1. Prepare all reagents before starting.
2. Add 100 µL of standards, controls, and samples to appropriate wells. Cover with adhesive strip and incubate at 37°C for 60 minutes.
3. Wash the plate 4 times manually or automatically.
4. Add 50 µL of Chromogen A and 50 µL of Chromogen B to each well. Incubate for 15 minutes at 37°C, protected from light.
5. Add 50 µL of Stop Solution to each well. The color will change from blue to yellow. If green or uneven color appears, gently tap the plate.
6. Measure OD at 450 nm within 15 minutes using a microplate reader.
**CALCULATION OF RESULTS**
1. Plot the average OD values of the standards against their concentrations to generate a standard curve.
2. Subtract the blank OD value from all measurements before interpretation.
3. Use graph paper or software to determine sample concentrations.
4. Intra-assay and inter-assay CVs are <15%.
5. Assay range: 1.56–50 ng/mL.
6. Sensitivity: <1.0 ng/mL.
7. Cross-reactivity: No significant cross-reaction with other cytokines.
8. Storage: 2–8°C (frequent use); -20°C (long-term storage).
**STORAGE & STABILITY**
- Store unopened kit at 2–8°C.
- Once opened, store unused strips in sealed bags with desiccant.
- Stability: 6 months at -20°C.
**NOTES**
This product is intended for research use only. Always follow proper safety protocols when handling biological materials. For detailed instructions, refer to the user manual provided.
Disposable forks are single-use utensils made of plastic or other materials that are designed to be thrown away after use. They are commonly used for eating take-out meals, at picnics, or for other outdoor events where it is not convenient or practical to use regular silverware. Disposable forks are lightweight, affordable, and easy to use, but they also contribute to plastic waste and environmental pollution. Some eco-friendly alternatives to disposable forks include biodegradable or Compostable Forks made from materials like cornstarch or bamboo.
Disposable Forks,Disposable Plastic Cutlery,Eco-Friendly Cutlery,Polypropylene Cutlery
Anhui Jianfeng Environmental Protection Technology Co., Ltd , https://www.ahbiocutleries.com