Human DKK1 ELISA Kit
2025-09-20 09:11:19
**Human DKK1 ELISA Kit – For the quantitative in vitro determination of Human Dickkopf 1 concentrations in serum, plasma, cerebrospinal fluid, tissue homogenate, and other body fluids. FOR LABORATORY RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES.**
Before using this product, please read this entire package insert carefully. This ELISA kit is designed for research purposes only and should not be used in diagnostic or clinical procedures.
**INTENDED USE AND TEST PRINCIPLE**
This DKK1 ELISA Kit is intended for laboratory research use only. The assay is based on the principle of a competitive immunoassay. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured. Standards are run simultaneously with the samples to generate a standard curve of Optical Density (OD) versus DKK1 concentration. The DKK1 concentration in the sample is then determined by comparing its OD value to the standard curve.
**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. 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. Avoid 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 granules are present.
**MATERIALS REQUIRED BUT NOT SUPPLIED**
1. Incubator set at 37°C
2. Microplate reader capable of measuring absorbance at 450 nm
3. Pipettes, disposable tips, and absorbent paper
4. Distilled or deionized water
**REAGENTS PROVIDED**
All reagents must be stored at 2–8°C. Check the expiration date on the label.
| Name | 96 Determinations | 48 Determinations |
|------|-------------------|-------------------|
| MicroELISA Stripplate | 12×8 strips | 12×4 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 |
**NOTES**
1. Standard concentrations: 160, 80, 40, 20, 10, 5 ng/mL.
2. If sample values exceed the highest standard, dilute with Sample Diluent and repeat the assay.
3. Allow all reagents and samples to reach room temperature (20–25°C) before use. Do not use water baths for thawing.
4. Do not use reagents past their expiration date.
5. Only use deionized or distilled water for dilutions.
6. Keep unused strips in the sealed bag with desiccant at 2–8°C.
7. Use fresh pipette tips for each transfer to prevent contamination.
8. Do not use any disposable knives that may have come into contact with rat blood, as there is no guarantee against infectious agents.
9. Liquid waste should be treated with sodium hypochlorite to a final concentration of 1.0% and left for at least 30 minutes before disposal.
10. Substrate solution is sensitive to contamination. Discard if it appears bluish.
11. Substrate B contains 20% acetone; keep away from heat or flame.
12. Bring all reagents to room temperature before use.
**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.
**ASSAY PROCEDURE**
1. Prepare all reagents before starting.
2. Add 100 µL of standards, samples, and blank to the microtiter plate.
3. Cover with adhesive strips and incubate for 60 minutes at 37°C.
4. Wash the plate 4 times manually or automatically.
- Manual: Aspirate, fill with Wash Solution, aspirate again. Repeat 4 times.
- Automated: Aspirate and wash four times. Adjust brush to remove as much liquid as possible. Fill 350 µL/well.
5. After washing, 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, protecting from light.
6. Add 50 µL of Stop Solution to each well. Read OD at 450 nm within 15 minutes.
**CALCULATION**
1. Plot average OD (450 nm) vs. standard concentrations.
2. Subtract blank OD from all values.
3. Generate a standard curve using graph paper or software.
4. Intersect the sample OD with the curve and read the corresponding concentration.
5. Intra-assay CV: <10%, Inter-assay range: 5–160 ng/mL.
6. Sensitivity: <1.0 ng/mL.
7. No cross-reactivity observed.
8. Storage: 2–8°C (frequent use), -20°C (long-term, up to 6 months).
**CAUTIONS AND WARNINGS**
- Always wear gloves and lab coat.
- Avoid direct contact with reagents.
- Follow proper waste disposal guidelines.
- Do not reuse reagents beyond the expiration date.
- Maintain consistent technique for accurate results.
**THIS KIT IS INTENDED FOR RESEARCH USE ONLY. DO NOT USE FOR DIAGNOSTIC PURPOSES.**
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