**Human Decorin ELISA Kit – For the quantitative in vitro determination of Human Decorin 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 for diagnostic or therapeutic applications.
**INTENDED USE AND TEST PRINCIPLE**
The Human Decorin ELISA Kit is intended for laboratory research use only. It utilizes a sandwich ELISA method to quantify human decorin levels in various biological samples. The colorimetric reaction is initiated by adding a chromogenic substrate, which changes from blue to yellow upon addition of the stop solution. The optical density (OD) is measured at 450 nm, and the concentration of decorin in the sample is determined by comparing the OD values to a standard curve generated from known concentrations.
**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. 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 for 30 minutes at 2–8°C. Store at -20°C. Avoid freeze-thaw cycles.
- **Cell culture supernatants, tissue homogenates, and other biological fluids**: Centrifuge to remove particulates and assay immediately or store at -20°C. Ensure no hemolysis or granules are present.
**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**
All reagents must be stored at 2–8°C. Check the expiration date on the label before use.
| Reagent | 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 |
**Note:**
- Standard concentrations: 200, 100, 50, 25, 12.5, 6.25 pg/mL
- If sample values exceed the highest standard, dilute with Sample Diluent and repeat the assay.
**PRECAUTIONS**
1. Do not mix reagents from different kit lots. All components are calibrated for optimal performance.
2. Allow all reagents and materials to reach room temperature (20–25°C) before use. Avoid using water baths for thawing.
3. Do not use reagents beyond their expiration date.
4. Use only deionized or distilled water for dilutions.
5. Keep microtiter plates in sealed bags until use. Store unused strips with desiccant at 2–8°C.
6. Use fresh pipette tips for each transfer to avoid contamination.
7. Handle all blood-derived products with care, as they may pose infectious risks.
8. Dispose of all samples and waste according to local regulations.
9. Liquid waste should be treated with sodium hypochlorite (1.0%) for at least 30 minutes before disposal.
10. Substrate solutions are sensitive to contamination. Avoid exposure to heat or flame.
11. Ensure all reagents are at room temperature before starting the assay.
**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 beginning. Run standards and samples in duplicate.
2. Add 50 μL of standard or sample to appropriate wells. Blank well contains no sample.
3. Add 100 μL of HRP-conjugate reagent to all wells.
4. Wash the plate four times with 1X Wash Solution. After the final wash, invert and blot dry.
5. Add 50 μL of Chromogen A and 50 μL of Chromogen B to each well. Incubate for 15 minutes at 37°C, away from light.
6. Add 50 μL of Stop Solution to each well. The color should turn yellow. If uneven, gently tap the plate.
7. Measure OD at 450 nm. Generate a standard curve by plotting average OD values against standard concentrations. Determine sample concentrations by interpolation.
**RESULT INTERPRETATION**
1. Calculate the mean OD for each standard and sample. Subtract blank OD from all readings.
2. Plot the standard curve and determine sample concentrations by matching OD values to the curve.
3. Intra-assay and inter-assay CV% are less than 15%.
4. Assay range: 6.25 pg/mL to 200 pg/mL.
5. Sensitivity: <1.0 pg/mL.
6. Cross-reactivity: No significant cross-reaction with other proteins.
7. Storage: 2–8°C (frequent use), or -20°C for up to six months.
**NOTES**
This kit is ideal for researchers studying extracellular matrix proteins, particularly in conditions such as fibrosis, wound healing, and connective tissue disorders. Always follow safety protocols and ensure proper handling of all reagents and samples.
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