**Human DcR3 ELISA Kit – For the Quantitative In Vitro Determination of Human Soluble Decoy Receptor (DcR3) in Serum, Plasma, Cerebrospinal Fluid, Tissue Homogenate, and Other Body Fluids**
*For Laboratory Research Use Only. Not for Diagnostic or Therapeutic Procedures.*
This ELISA kit is designed for the quantitative measurement of soluble Decoy Receptor 3 (DcR3) in various biological samples. The assay is based on the enzyme-linked immunosorbent assay (ELISA) principle, where a standard curve is generated by measuring optical density (OD) at 450 nm. The concentration of DcR3 in unknown samples is determined by comparing their OD values to the standard curve.
**Intended Use:**
The Human DcR3 ELISA Kit is intended for research purposes only and should not be used in diagnostic or therapeutic procedures. It provides a reliable method for quantifying DcR3 levels in different biological matrices, such as serum, plasma, cerebrospinal fluid, tissue homogenates, and other body fluids.
**Sample Collection and Storage:**
- **Serum:** Collect using a serum separator tube. Allow clotting for 2 hours at room temperature or overnight at 4°C before centrifuging at 2000×g for 20 minutes. Remove serum and assay immediately or store at -20°C. Avoid repeated freeze-thaw cycles.
- **Plasma:** Collect with heparin as an anticoagulant. Centrifuge within 30 minutes at 2000×g, 2–8°C. Store at -20°C.
- **Cell Culture Supernatants, Tissue Homogenates, and Other Fluids:** Centrifuge to remove particulates and assay immediately or store at -20°C. Ensure no hemolysis or granules are present in the samples.
**Materials Required (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 (Stored at 2–8°C):**
- Microtiter Strip Plate: 12×8 strips (96 determinations), 12×4 strips (48 determinations)
- Standard (6 vials): 0.5 mL/vial
- 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 units
- User Manual: 1 copy
- Sealed Bags: 1 per kit
**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 past their expiration date.
4. Use only deionized or distilled water for dilutions.
5. Keep unused microtiter plate strips in sealed bags with desiccant at 2–8°C.
6. Use fresh pipette tips for each transfer to prevent contamination.
7. Treat all biological materials as potentially infectious. Follow proper biosafety protocols.
8. Dispose of all samples and waste according to local regulations. Allow liquid to stand for 30 minutes to inactivate viruses before disposal.
9. Avoid contact with skin or eyes. Wear appropriate personal protective equipment (PPE).
10. Substrate solutions may be easily contaminated. Handle with care and protect from light.
**Reagent Preparation and Storage:**
- **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 and add 50 µL of standard or sample to appropriate wells (excluding blank well).
2. Cover with adhesive strip and incubate for 60 minutes at 37°C.
3. Wash the microtiter plate 4 times manually or using automated washing system.
4. Add 50 µL of Chromogen A and 50 µL of Chromogen B to each well. Incubate for 15 minutes at 37°C, protecting from light.
5. Add 50 µL of Stop Solution to each well. Read OD at 450 nm using a microplate reader.
6. Generate a standard curve by plotting average OD values against DcR3 concentrations. Calculate sample concentrations by comparing OD to the curve.
**Performance Characteristics:**
- **Intra-assay CV (%) and Inter-assay CV (%):** <15%
- **Assay Range:** 3.75 pg/mL – 120 pg/mL
- **Sensitivity:** <1.0 pg/mL
- **Cross-reactivity:** No significant cross-reactivity observed with other proteins.
- **Storage:** 2–8°C (frequent use); 6 months at -20°C.
**Note:** If sample values exceed the highest standard, dilute with Sample Diluent and repeat the assay. Always read the entire package insert carefully before use.
**Important:** This kit is for laboratory research only. Not for human diagnostic use.
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