**Human Scd163 ELISA Kit – For the Quantitative In Vitro Determination of Human Soluble CD163 Concentrations in Serum, Plasma, Cerebrospinal Fluid, Tissue Homogenate, and Other Body Fluids**
*For Laboratory Research Use Only. Not for Diagnostic or Therapeutic Procedures.*
This kit is designed for the quantitative measurement of soluble Cluster of Differentiation 163 (Scd163) in various biological samples. The assay is based on the enzyme-linked immunosorbent assay (ELISA) principle, using a competitive binding format. The Stop Solution changes the color from blue to yellow, and the intensity of the color is directly proportional to the concentration of Scd163 in the sample. A set of calibration standards is provided to generate a standard curve, allowing for accurate quantification of Scd163 levels in unknown samples.
**Intended Use:**
The Human Scd163 ELISA Kit is intended for laboratory research purposes only and should not be used for diagnostic or therapeutic procedures. It is suitable for measuring Scd163 concentrations in serum, plasma, cerebrospinal fluid, tissue homogenates, and other body fluids.
**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. Remove serum and analyze immediately or aliquot and store at -20°C. Avoid repeated freeze-thaw cycles.
- **Plasma:** Collect using heparin as an anticoagulant. Centrifuge within 30 minutes at 2000×g (2–8°C). Store at -20°C and avoid freeze-thaw cycles.
- **Cell culture supernatants, tissue homogenates, and other biological fluids:** Centrifuge to remove particulates and analyze immediately or store at -20°C. Thaw only once.
*Note: Ensure proper centrifugation and avoid hemolysis or particulate contamination.*
**Materials Required but Not Supplied:**
1. Incubator 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):**
- 96-well microplate (12×8 strips)
- 6 vials of standard (0.5 mL/vial)
- Sample diluent (6.0 mL / 3.0 mL)
- HRP-conjugated reagent (10.0 mL / 5.0 mL)
- 20X Wash solution (25 mL / 15 mL)
- Chromogen A (6.0 mL / 3.0 mL)
- Chromogen B (6.0 mL / 3.0 mL)
- Stop solution (6.0 mL / 3.0 mL)
- Closure plate membranes (2 units)
- User manual and sealed bags
**Standard Concentrations:**
160, 80, 40, 20, 10, 5 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 and materials to reach room temperature (20–25°C) before use. Do not use water baths for thawing.
3. Do not use reagents beyond their expiration date.
4. Store unused strips in their pouch with desiccant at 2–8°C.
5. Wear gloves during the procedure. Handle all blood-derived samples as potentially infectious.
6. Dispose of waste properly after inactivation (allow standing for 30 minutes to inactivate viruses).
7. Keep chromogen B away from heat or flame (contains 20% acetone).
**Reagent Preparation:**
- Wash Solution (1X): Dilute 1 volume of 20X wash solution with 19 volumes of distilled or deionized water. Store at 2–8°C for 1 month.
**Assay Procedure:**
1. Prepare all reagents before starting. Run standards and samples in duplicate.
2. Add 50 µL of standard or sample to appropriate wells (blank well receives nothing).
3. Add 100 µL of HRP-conjugate reagent to all wells except the blank. Cover with adhesive strip and incubate for 60 minutes at 37°C.
4. Wash 4 times manually or automatically.
5. 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.
6. Add 50 µL of stop solution to each well. Color changes from blue to yellow.
7. Read OD at 450 nm. Plot average OD values of standards to create a standard curve. Subtract blank OD from all readings.
**Performance Characteristics:**
- Intra-assay CV < 15%, Inter-assay CV < 15%
- Assay range: 5–160 ng/mL
- Sensitivity: <1.0 ng/mL
- No cross-reactivity or interference observed
**Storage:**
- 2–8°C (frequent use)
- -20°C (long-term storage) for up to 6 months
**Important Notes:**
- Always read the entire user manual before beginning the assay.
- Each operator should generate their own standard curve.
- Variations in technique, timing, or temperature may affect results.
**Quote:**
"Accuracy and consistency depend on strict adherence to the protocol."
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