**Human WISP1 ELISA Kit – For the Quantitative In Vitro Determination of Human WNT1 Inducible Signaling Pathway Protein 1 in Serum, Plasma, Cerebrospinal Fluid, Tissue Homogenate, and Other Body Fluids**
**For Laboratory Research Use Only. Not for Diagnostic or Therapeutic Procedures.**
**INTENDED USE AND TEST PRINCIPLE**
This WISP1 ELISA Kit is designed for research purposes only and is not intended for use in diagnostic or therapeutic procedures. The kit utilizes a sandwich ELISA method to quantitatively measure Human WISP1 levels in various biological samples. The reaction involves binding of WISP1 to specific antibodies coated on microtiter strips, followed by detection with a horseradish peroxidase (HRP)-conjugated secondary antibody. A chromogenic substrate is added, resulting in a color change that is proportional to the WISP1 concentration. The Stop Solution terminates the reaction, and optical density (OD) is measured at 450 nm. By comparing sample OD values to a standard curve, the exact WISP1 concentration can be determined.
**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 centrifuging at 2000×g for 20 minutes. Remove serum and assay 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. Avoid freeze-thaw cycles.
- **Cell Culture Supernatants, Tissue Homogenates, and Other Biological Fluids**: Centrifuge to remove particulates and assay immediately, or store at -20°C. Thaw only once.
*Note: Ensure samples are properly centrifuged, and avoid hemolysis or contamination.*
**MATERIALS REQUIRED BUT NOT SUPPLIED**
1. 37°C incubator
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**
All reagents should be stored at 2–8°C. Check expiration date on label.
- MicroELISA Stripplate: 12 × 8 strips / 12 × 4 strips
- 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 / 2
- User Manual: 1 / 1
- Sealed Bags: 1 / 1
**PRECAUTIONS**
1. Do not substitute reagents from different kit lots. Standards, conjugates, and plates are matched for optimal performance.
2. Allow all reagents 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 sealed pouches with desiccant at 2–8°C.
5. Wear disposable gloves during handling. Treat all samples as potentially infectious.
6. Dispose of waste after inactivation with 1% sodium hypochlorite for at least 30 minutes.
7. Keep Chromogen Solutions A and B away from heat and flame.
8. Follow good laboratory practices at all times.
**REAGENT PREPARATION AND STORAGE**
- **Wash Solution (1X)**: Dilute 1 volume of 20X Wash Solution with 19 volumes of distilled or deionized water. Store at 2–8°C for up to 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 no addition.
3. Add 100 µL HRP-conjugate reagent to all wells except blank. Cover with adhesive strip and incubate for 60 minutes at 37°C.
4. Wash plate 4 times manually or automatically.
5. Add 50 µL each of Chromogen A and B. Incubate 15 minutes at 37°C, protected from light.
6. Add 50 µL Stop Solution. Mix gently if color is uneven.
7. Read OD at 450 nm. Plot standard curve (OD vs. concentration). Determine sample concentrations by comparison.
**QUALITY CONTROL**
- Intra-assay and Inter-assay CV <15%.
- Assay range: 31.2 pg/mL – 1000 pg/mL.
- Sensitivity: <10 pg/mL.
- Cross-reactivity: No significant cross-reaction with other proteins.
- Storage: 2–8°C (frequent use); -20°C for long-term (up to 6 months).
**NOTES**
Always read the full instructions before performing the assay. Maintain accurate records and follow all safety guidelines. This kit is not intended for clinical use.
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