**Human LPA ELISA Kit – For the Quantitative In Vitro Determination of Human Lysophosphatidic Acid (LPA) Concentrations in Serum, Plasma, Cerebrospinal Fluid, Tissue Homogenate, and Other Body Fluids**
*For Laboratory Research Use Only. Not for Diagnostic or Therapeutic Procedures.*
Before using this product, please read this entire package insert carefully. This ELISA kit is designed for research purposes only and should not be used in diagnostic or clinical settings.
---
### **INTENDED USE AND TEST PRINCIPLE**
This Human LPA ELISA Kit is intended for laboratory research use only and is not suitable for diagnostic or therapeutic procedures. The method is based on a competitive immunoassay. The calibration standards are run alongside the samples to generate a standard curve by plotting optical density (OD) values against known LPA concentrations. The LPA concentration in each sample is then determined by comparing its OD value to the standard curve.
---
### **SAMPLE COLLECTION AND STORAGE**
- **Serum**: Collect using 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. 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). Store at -20°C. Avoid multiple freeze-thaw cycles.
- **Cell Culture Supernatants, Tissue Homogenates, and Other Biological Fluids**: Remove particulates by centrifugation. Assay immediately or store at -20°C. Ensure no hemolysis or contamination occurs during preparation.
---
### **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**
| Reagent | 96 Determinations | 48 Determinations |
|---------|-------------------|-------------------|
| MicroELISA Strip Plate | 12×8 strips | 12×4 strips |
| Standard (6 Vials) | 0.5 ml/vial | 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 |
**Note:** Standard concentrations: 3200, 1600, 800, 400, 200, 100 pmol/mL. If sample values exceed the highest standard, dilute with Sample Diluent and retest.
---
### **PRECAUTIONS**
1. Do not mix reagents from different kit lots. All components are optimized for 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. Use only deionized or distilled water for dilutions.
5. Keep unused microplate strips in sealed bags with desiccant at 2–8°C.
6. Use fresh pipette tips for each transfer to prevent cross-contamination.
7. Treat all biological materials as potentially infectious. Follow standard lab safety protocols.
8. Dispose of all waste following local regulations. Allow waste to stand for at least 30 minutes to inactivate viruses.
9. Substrate solutions are sensitive; avoid contamination. Store away from heat and flame.
10. Always allow reagents to equilibrate to room temperature before use.
---
### **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 1 month.
---
### **ASSAY PROCEDURE**
1. Prepare all reagents before starting. Add 50 µL of standard or sample to appropriate wells. Cover with adhesive strip and incubate for 60 minutes at 37°C.
2. Wash the microtiter plate 4 times using either manual or automated washing methods. After final wash, blot dry gently.
3. Add 50 µL of Chromogen Solution A and 50 µL of Chromogen Solution B to each well. Mix gently and incubate for 15 minutes at 37°C, protecting from light.
4. Add 50 µL of Stop Solution to each well. Read OD at 450 nm using a microplate reader.
---
### **DATA ANALYSIS**
1. Plot average OD values (450 nm) of standards vs. concentrations on graph paper or software.
2. Subtract blank OD from all readings before interpretation.
3. Determine LPA concentration by locating the sample OD on the Y-axis and drawing a line to intersect the standard curve. Then draw a vertical line to the X-axis to find the corresponding concentration.
4. Intra-assay and inter-assay CV% < 15%.
5. Assay range: 100–3200 pmol/mL.
6. Sensitivity: <100 pmol/mL. No significant cross-reactivity observed.
---
### **STORAGE**
- Store at 2–8°C for frequent use. For long-term storage, keep at -20°C for up to 6 months.
---
**Important:** Always follow the manufacturer's instructions and maintain proper lab practices. This product is not intended for human diagnosis or treatment.
Make Up,Whitening Moisturizing Foundation,Waterproof Whiten Skin Bb Cream,Air Cushion Bb Cream
RYEOGLOBAL Co,.LTD , https://www.ryeolab.com