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**Human Corin ELISA Kit – For the quantitative in vitro determination of Human Corin concentrations in serum, plasma, cerebrospinal fluid, tissue homogenate, and other body fluids. Intended for laboratory research use only. NOT FOR USE IN DIAGNOSTIC PROCEDURES.**
Before using this product, please read the entire package insert carefully.
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### **INTENDED USE AND TEST PRINCIPLE**
This Human Corin ELISA Kit is specifically designed for research purposes and is not intended for diagnostic or therapeutic use. The kit utilizes a sandwich ELISA technique to quantitatively measure Human Corin levels in various biological samples. The color change from blue to yellow upon addition of the Stop Solution indicates the reaction completion. A standard curve is generated using known concentrations of Corin, allowing for accurate quantification of unknown samples by comparing their optical density (OD) values to the standard curve.
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### **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 centrifugation 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 of collection at 2000×g for 30 minutes at 2–8°C. Store at -20°C and avoid repeated freeze-thaw cycles.
- **Cell culture supernatants, tissue homogenates, and other biological fluids**: Centrifuge to remove particulates. Assay immediately or store at -20°C. Ensure no hemolysis or granulation occurs.
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### **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
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### **REAGENTS PROVIDED**
All reagents should be stored at 2–8°C. Check the expiration date on the label before use.
| Reagent Name | 96 Determinations | 48 Determinations |
|-------------------------------|-------------------|-------------------|
| MicroELISA Stripplate | 12×8 strips | 12×4 strips |
| Standard (6 Vial) | 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: 50, 25, 12.5, 6.25, 3.12, 1.56 ng/mL
- If sample values exceed the highest standard, dilute with Sample Diluent and repeat the assay.
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### **PRECAUTIONS**
1. Do not mix reagents from different kit lots. All components are calibrated for optimal performance.
2. Allow all reagents to reach room temperature (20–25°C) before use.
3. Do not thaw samples or reagents in water baths.
4. Do not use reagents beyond their expiration date.
5. Use only deionized or distilled water for reagent dilutions.
6. Keep microtiter plates in sealed bags until needed. Unused strips should be stored with desiccant at 2–8°C.
7. Use fresh pipette tips for each transfer to prevent cross-contamination.
8. Wear gloves during the procedure to minimize contamination risk.
9. Dispose of waste after at least 30 minutes of inactivation to neutralize potential viruses.
10. Substrate solution may be easily contaminated. Discard if it appears bluish.
11. Chromogen B contains 20% acetone; keep away from heat and flame.
12. Always read the instructions thoroughly before starting the test.
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### **REAGENT PREPARATION AND STORAGE**
- **Wash Solution (1X)**: Mix 1 volume of 20X Wash Solution with 19 volumes of deionized or distilled water. Store at 2–8°C for up to one month.
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### **ASSAY PROCEDURE**
1. Prepare all reagents before beginning.
2. Add 100 μL of standards, samples, and blank to the microplate. Cover with adhesive strips and incubate for 60 minutes at 37°C.
3. Wash the plate 4 times manually or using automated washing.
- Manual: Aspirate, fill with 1X Wash Solution, aspirate again. Repeat 4 times. Dry by blotting.
- Automated: Aspirate, wash 4 times, adjust brush and fill volume to 350 μL/well.
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. The color should turn from blue to yellow. Gently tap if uneven.
6. Read OD at 450 nm within 15 minutes.
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### **CALCULATION OF RESULTS**
1. Plot average OD values of standards against their concentrations on graph paper or software.
2. Subtract blank OD from all readings before interpretation.
3. Locate the sample OD on the Y-axis, draw a horizontal line to the curve, then a vertical line to the X-axis to determine concentration.
4. Each user must generate their own standard curve due to possible variations in technique, timing, or kit age.
5. Intra-assay and inter-assay CVs are less than 15%.
6. Assay range: 1.56 ng/mL – 50 ng/mL.
7. Sensitivity: <1.0 ng/mL.
8. Cross-reactivity: No significant cross-reaction with other proteins.
9. Storage: 2–8°C (frequent use), or -20°C for long-term storage (up to 6 months).
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**Please ensure all steps are followed accurately to obtain reliable results.**
**For technical support, contact the manufacturer.**