**Human MIP-1δ ELISA Kit – For the quantitative in vitro determination of Human Macrophage Inflammatory Protein 1δ concentrations in serum, plasma, cerebrospinal fluid, tissue homogenate, and other body fluids. Intended for laboratory research use only. NOT FOR USE IN DIAGNOSTIC PROCEDURES.**
This ELISA kit is designed for the accurate measurement of MIP-1δ levels in various biological samples. The principle of the assay is based on a sandwich ELISA format. The colorimetric reaction is initiated by adding a chromogenic substrate, which changes color in the presence of HRP enzyme. The reaction is stopped with a stop solution, turning the color from blue to yellow. The optical density (OD) is measured at 450 nm using a microplate reader.
To determine the concentration of MIP-1δ in your samples, this kit includes a set of standard solutions with known concentrations. These standards are run alongside your samples, allowing you to create a standard curve. By comparing the OD values of your samples to the standard curve, you can accurately calculate the MIP-1δ concentration.
**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 (20 minutes at ~2000×g). Remove the serum and analyze immediately or store at -20°C. Avoid repeated freeze-thaw cycles.
- **Plasma:** Collect using heparin as an anticoagulant. Centrifuge within 30 minutes at 2–8°C for 30 minutes at 2000×g. Store at -20°C. Avoid repeated freeze-thaw.
- **Cell culture supernatants, tissue homogenates, and other body fluids:** Centrifuge to remove particulates and analyze immediately or store at -20°C. Ensure no hemolysis or granules are present.
**Materials Required (Not Supplied):**
- 37°C incubator
- 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):**
- Microtiter strip plate (12×8 or 12×4 strips)
- Standards (6 vials, 0.5 ml/vial)
- Sample diluent (6.0 ml or 3.0 ml)
- HRP-conjugated reagent (10.0 ml or 5.0 ml)
- 20X Wash solution (25 ml or 15 ml)
- Chromogen A (6.0 ml or 3.0 ml)
- Chromogen B (6.0 ml or 3.0 ml)
- Stop solution (6.0 ml or 3.0 ml)
- Closure plate membrane (2 pieces)
- User manual and sealed bags
**Precautions:**
- Do not mix reagents from different kits. All components are matched for optimal performance.
- Bring all reagents to room temperature (20–25°C) before use.
- Do not use beyond the expiration date.
- Store unused strips in the provided desiccant bag at 2–8°C.
- Always wear gloves when handling biological samples.
- Dispose of waste properly after adding sodium hypochlorite to inactivate pathogens.
- Handle chromogen solutions carefully, as they contain acetone and should be kept away from heat or flame.
**Assay Procedure:**
1. Prepare all reagents before starting.
2. Add 50 μL of standard or sample to appropriate wells (duplicate).
3. Add 100 μL of HRP-conjugate reagent to each well except the blank. Incubate for 60 minutes at 37°C.
4. Wash the plate 4 times manually or automatically.
5. Add 50 μL of Chromogen A and 50 μL of Chromogen B to each well. Incubate for 15 minutes at 37°C, protected from light.
6. Add 50 μL of Stop Solution. Mix gently if color change is uneven.
7. Measure OD at 450 nm. Plot the standard curve and calculate sample concentrations accordingly.
**Performance Characteristics:**
- Sensitivity: <10 pg/mL
- Assay range: 15–480 pg/mL
- Intra-assay and inter-assay CV <15%
- No significant cross-reactivity with other proteins
**Storage:**
- Store at 2–8°C for frequent use; up to 6 months at -20°C.
**Note:** This kit is intended for research purposes only and should not be used for diagnostic or therapeutic applications. Always follow good laboratory practices and safety protocols when handling biological materials.
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