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Human hs-CRP ELISA Kit
**Human hs-CRP ELISA Kit – For the Quantitative In Vitro Determination of Human High-Sensitivity C-Reactive Protein in Serum, Plasma, Cerebrospinal Fluid, Tissue Homogenate, and Other Body Fluids**
*For Laboratory Research Use Only. Not for Diagnostic or Therapeutic Procedures.*
This high-sensitivity C-Reactive Protein (hs-CRP) ELISA kit is designed for the quantitative measurement of hs-CRP concentrations in various biological samples. The assay is based on a sandwich immunoassay principle, where the colorimetric reaction is detected at 450 nm using a spectrophotometer. A standard curve is generated using a series of calibration standards, allowing for accurate quantification of hs-CRP levels in test samples.
**Intended Use**
The kit is intended for laboratory research purposes only and is not suitable for diagnostic or clinical use. It provides a reliable method to measure low levels of CRP, which is an important biomarker for inflammation and cardiovascular risk assessment.
**Sample Collection and Storage**
- **Serum**: Collect using a serum separator tube. Allow clotting for 2 hours at room temperature or overnight at 4°C. Centrifuge at 2000×g for 20 minutes. Store at -20°C after aliquoting. Avoid repeated freeze-thaw cycles.
- **Plasma**: Use heparin as an anticoagulant. Centrifuge within 30 minutes at 2000×g (2–8°C). Avoid freeze-thaw.
- **Tissue Homogenate, Cell Culture Supernatant, and Other Body Fluids**: Centrifuge to remove particulates. Assay immediately or store at -20°C. Thaw only once.
**Materials Required but Not Supplied**
1. Incubator 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**
- Microtiter Strip Plate (12 x 8 strips or 12 x 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)
**Standard Concentrations**
400, 200, 100, 50, 25, 12.5 ng/mL
**Precautions**
- Do not mix reagents from different kit lots.
- Bring all reagents to room temperature before use.
- Do not use beyond expiration date.
- Store unused strips with desiccant at 2–8°C.
- Handle plasma as potentially hazardous. Wear gloves during the procedure.
- Discard waste after 30 minutes of inactivation.
- Avoid contact with acid and sodium hypochlorite.
- Substrate B contains 20% acetone; keep away from heat and flame.
**Reagent Preparation**
- Wash Solution (1X): Dilute 1 volume of 20X Wash Solution with 19 volumes of distilled water. Stable for 1 month at 2–8°C.
**Assay Procedure**
1. Prepare all reagents. Add standards and samples in duplicate.
2. Add 50 µL of standard or sample to each well. Blank well: no addition.
3. Add 100 µL of HRP-conjugate reagent to all wells except blank. Cover and incubate for 60 minutes at 37°C.
4. Wash plate 4 times manually or automatically.
5. Add 50 µL of Chromogen A and B to each well. Incubate for 15 minutes at 37°C, protected from light.
6. Add 50 µL of Stop Solution. Color changes from blue to yellow. Mix gently if needed.
7. Read OD at 450 nm. Plot standard curve and calculate sample concentrations.
**Performance Characteristics**
- Sensitivity: <10 ng/mL
- Range: 12.5–400 ng/mL
- Intra-assay and Inter-assay CV <15%
- No significant cross-reactivity observed
**Storage**
- 2–8°C for frequent use
- -20°C for long-term storage (up to 6 months)
**Note:** Always follow proper safety protocols and dispose of waste according to local regulations. Each user should generate their own standard curve for optimal accuracy.