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Human CEACAM1 ELISA Kit
**Human CEACAM1 ELISA Kit – For Quantitative In Vitro Determination of Human CEACAM1 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 kit, please read the entire package insert carefully. This ELISA (Enzyme-Linked Immunosorbent Assay) is designed for the quantitative measurement of Human CEACAM1 in various biological samples. The principle of the assay involves binding CEACAM1 to a microtiter plate coated with specific antibodies. After incubation with a horseradish peroxidase (HRP)-conjugated secondary antibody, a chromogenic substrate is added, resulting in a color change that is proportional to the concentration of CEACAM1. The reaction is stopped, and absorbance is measured at 450 nm. A standard curve is generated from known concentrations, allowing for accurate quantification of unknown samples.
**Sample Collection and Storage:**
- **Serum:** Use a serum separator tube. Allow clotting for 2 hours at room temperature or overnight at 4°C. Centrifuge 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. Store at -20°C.
- **Cell Culture Supernatant, Tissue Homogenate, and Other Biological Fluids:** Centrifuge to remove particulates. Assay immediately or store at -20°C. Ensure no hemolysis or granules are present.
**Materials Required but Not Supplied:**
- Incubator at 37°C
- Microplate reader capable of measuring absorbance at 450 nm
- Precision pipettes, disposable tips, and absorbent paper
- Distilled or deionized water
**Reagents Provided (Stored at 2–8°C):**
- MicroELISA Strip Plate (12×8 or 12×4 strips)
- Standard (6 vials, 0.5 ml/vial)
- Sample Diluent (6.0 ml or 3.0 ml)
- HRP-Conjugate Reagent (10.0 ml or 5.0 ml)
- 20X Wash Solution (25 ml or 15 ml)
- Chromogen Solution A (6.0 ml or 3.0 ml)
- Chromogen Solution B (6.0 ml or 3.0 ml)
- Stop Solution (6.0 ml or 3.0 ml)
- Closure Plate Membrane (2 pieces)
- User Manual (1 copy)
- Sealed Bags (1 bag)
**Precautions:**
- Never substitute reagents between different kit lots.
- Allow all reagents to reach room temperature before use.
- Do not use beyond the expiration date.
- Use only deionized or distilled water.
- Keep microtiter plates in sealed bags until needed.
- Use fresh pipette tips to prevent cross-contamination.
- All blood-derived samples should be treated as potentially infectious.
- Dispose of waste properly after inactivation for at least 30 minutes.
- Avoid contact with heat or flame when handling Substrate B, which contains 20% acetone.
**Reagent Preparation:**
- Wash Solution (1X): Dilute 1 volume of 20X Wash Solution with 19 volumes of water. Store at 2–8°C for up to one month.
**Assay Procedure:**
1. Prepare all reagents and set up the microplate.
2. Add 50 µL of standard or sample to each well (duplicate).
3. Add 100 µL of HRP-conjugate reagent to all wells except the blank.
4. Incubate for 60 minutes at 37°C.
5. Wash the plate four times (manual or automated).
6. Add 50 µL of Chromogen A and 50 µL of Chromogen B. Incubate for 15 minutes at 37°C, protected from light.
7. Add 50 µL of Stop Solution to each well. The color will turn yellow.
8. Read OD at 450 nm. Generate a standard curve by plotting OD vs. CEACAM1 concentration.
**Interpretation of Results:**
- Calculate the mean OD for each standard and sample.
- Subtract blank OD values before plotting.
- Use the standard curve to determine CEACAM1 concentration in unknown samples.
- Intra-assay and inter-assay CV values are less than 15%.
- Detection range: 125 pg/mL – 4000 pg/mL.
- Sensitivity: <100 pg/mL.
- No significant cross-reactivity observed.
**Storage:**
- Store unused kit components at 2–8°C for frequent use, or at -20°C for long-term storage (up to 6 months).
Always follow proper safety protocols and dispose of all materials according to local regulations. This kit is intended for research purposes only and should not be used for clinical diagnosis.