Human DKK1 ELISA Kit

**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 PROCEDURES.** Before using this product, please read the entire package insert carefully. This kit is designed for research purposes only and should not be used in diagnostic or therapeutic applications. **INTENDED USE AND TEST PRINCIPLE** The Human DKK1 ELISA Kit is intended for laboratory research use only. It utilizes a sandwich ELISA method to quantitatively measure DKK1 levels in biological samples. The reaction involves binding of DKK1 to specific antibodies immobilized on microtiter strips. A chromogenic substrate is then added, producing a color change that correlates with DKK1 concentration. The optical density (OD) is measured at 450 nm, and results are calculated by comparing sample OD values to a 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 of collection at 2000×g for 30 minutes at 2–8°C. Store at -20°C. Avoid freeze-thaw cycles. - **Cell culture supernatants, tissue homogenates, and other body fluids**: Centrifuge to remove particulates. Assay immediately or store at -20°C. Ensure no hemolysis or granulation occurs. **MATERIALS REQUIRED BUT NOT SUPPLIED** - 37°C incubator - Microplate reader capable of measuring absorbance at 450 nm - Pipettes, disposable tips, and absorbent paper - Distilled or deionized water **REAGENTS PROVIDED** All reagents must be stored at 2–8°C. Check the expiration date on the label. | Reagent Name | 96 Determinations | 48 Determinations | |----------------------------|-------------------|-------------------| | MicroELISA Strip Plate | 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. 3. Allow all reagents and samples to reach room temperature (20–25°C) before use. Do not thaw using water baths. 4. Do not use reagents beyond 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 cross-contamination. 8. Do not use disposable knives for the assay due to potential contamination risks. 9. Liquid waste must be treated with 1.0% sodium hypochlorite for 30 minutes before disposal. 10. Substrate solutions should be checked for discoloration before use. 11. Substrate B contains 20% acetone; keep away from heat and flame. 12. Allow all reagents to reach room temperature before starting the assay. **REAGENT PREPARATION** - **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 and set up the microtiter plate. 2. Add standards, samples, and blank to the wells. Cover with adhesive strips and incubate for 60 minutes at 37°C. 3. Wash the plate 4 times manually or automatically. 4. 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. 5. Add 50 µL of Stop Solution to each well. Read OD at 450 nm within 15 minutes. **CALCULATION** Plot average OD values (450 nm) against standard concentrations. Subtract blank OD from all readings. Use graph paper or software to generate the standard curve. Determine DKK1 concentration by interpolating sample OD values on the curve. **SPECIFICATIONS** - Sensitivity: <1.0 ng/mL - Intra-assay CV: <10% - Inter-assay range: 5–160 ng/mL - Cross-reactivity: No significant cross-reactivity observed - Storage: 2–8°C (for frequent use); -20°C for long-term storage (up to 6 months) **CAUTIONS** Results may vary based on operator technique, incubation time, and reagent stability. Each user should establish their own standard curve. Always follow safety protocols when handling reagents and waste. **FOR RESEARCH USE ONLY**

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