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Human CEACAM1 ELISA Kit
**Human CEACAM1 ELISA Kit – For the Quantitative In Vitro Determination of Human CEACAM1 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 the entire package insert carefully. This ELISA kit is designed for research purposes only and is not intended for clinical diagnosis or treatment.
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### **INTENDED USE AND TEST PRINCIPLE**
The Human CEACAM1 ELISA Kit is specifically designed for the quantitative determination of CEACAM1 levels in biological samples such as serum, plasma, cerebrospinal fluid, tissue homogenates, and other body fluids. The test is based on the enzyme-linked immunosorbent assay (ELISA) principle, where a colorimetric reaction occurs in proportion to the concentration of CEACAM1 present in the sample.
A standard curve is generated by measuring the optical density (OD) of known CEACAM1 concentrations. The OD values of the unknown samples are then compared to this curve to determine their CEACAM1 levels.
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### **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 the 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 of collection at 2000×g for 30 minutes at 2–8°C. Store at -20°C and avoid repeated freezing.
- **Cell Culture Supernatants, Tissue Homogenates, and Other Biological Fluids:** Centrifuge to remove particulates. Assay immediately or store at -20°C. Ensure no hemolysis or granules are present in the samples.
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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 (Stored at 2–8°C)**
| Reagent Name | 96 Determinations | 48 Determinations |
|--------------|-------------------|-------------------|
| Micro ELISA Strip Plate | 12 x 8 strips | 12 x 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: 4000, 2000, 1000, 500, 250, 125 pg/ml
- If sample values exceed the highest standard, dilute with Sample Diluent and repeat the assay.
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### **PRECAUTIONS**
1. Do not substitute 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 any reagent past its expiration date.
4. Only use deionized or distilled water.
5. Keep microtiter plates in their sealed bags until needed. Unused strips should be stored with desiccant at 2–8°C.
6. Use fresh disposable pipette tips for each transfer to prevent contamination.
7. Wear disposable gloves and lab coats during the procedure. Treat all samples as potentially infectious.
8. Dispose of waste properly after inactivation. Allow waste to stand for at least 30 minutes to inactivate viruses.
9. Substrate solutions are sensitive to contamination. Handle with care.
10. Substrate B contains 20% acetone—keep away from heat and flame.
11. Always bring reagents to room temperature before use.
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### **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.
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### **ASSAY PROCEDURE**
1. Prepare all reagents and standards before starting.
2. Add 50 μl of standard or sample to appropriate wells. Include a blank well without any addition.
3. Add 100 μl of HRP-conjugate reagent to all wells except the blank. Cover with an adhesive strip and incubate for 60 minutes at 37°C.
4. Wash the microtiter plate 4 times.
- *Manual Washing:* Aspirate, fill with 1X Wash Solution, and repeat 4 times. Blot dry.
- *Automated Washing:* Use 1X wash buffer. Adjust brush to aspirate as much liquid as possible. Set fill volume to 350 μL/well.
5. Add 50 μl of Chromogen A and 50 μl of Chromogen B to each well. Mix gently and incubate for 15 minutes at 37°C, protected from light.
6. Add 50 μl of Stop Solution to each well. The color should change from blue to yellow. If green or uneven color appears, gently tap the plate.
7. Measure OD at 450 nm using a microplate reader.
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### **RESULT INTERPRETATION**
- Plot the average OD values of the standards against their concentrations to create a standard curve.
- Subtract the blank OD from all measurements before interpretation.
- Locate the sample OD on the Y-axis and draw a horizontal line to intersect the standard curve. From there, draw a vertical line to the X-axis to determine the CEACAM1 concentration.
- Each user should generate their own standard curve.
- Intra-assay and inter-assay CV% are less than 15%.
- Assay range: 125 pg/ml – 4000 pg/ml
- Sensitivity: <100 pg/ml
- Cross-reactivity: No significant cross-reaction observed.
- Storage: 2–8°C (frequent use); -20°C (long-term storage).
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**Important:** This kit is for research use only. Do not use it for diagnostic or therapeutic purposes. Follow all safety and handling instructions carefully.