**Rat Neutrophil Elastase (NE) ELISA Kit Instructions for Use**
This kit is intended for research purposes only and is designed to quantify neutrophil elastase (NE) in rat serum, plasma, and other biological fluids. The assay is based on the sandwich ELISA technique, which allows for accurate and specific detection of NE levels in samples.
**Principle of the Assay**
The ELISA procedure involves immobilizing a purified anti-NE antibody onto a microplate. After incubation with the sample, a horseradish peroxidase (HRP)-labeled anti-NE antibody binds to the captured NE, forming an immune complex. Following washing steps, the substrate TMB is added, resulting in a color change that is proportional to the NE concentration. The reaction is stopped, and the absorbance is measured at 450 nm using a microplate reader. A standard curve is then used to calculate the NE concentration in the samples.
**Kit Components**
The kit includes all necessary reagents and materials for performing the ELISA. It comes in 48-well or 96-well configurations, with the following components:
- Coated Microplate (1 × 48 or 1 × 96)
- Standard Solutions (27 ng/mL, 0.5 mL × 1 bottle)
- Standard Diluent (1.5 mL × 1 bottle)
- Enzyme-Labeled Reagent (3 mL × 1 bottle)
- Substrate A and B (3 mL × 1 bottle each)
- Wash Buffer (20× concentrated, 20 mL × 20 times or 30 times)
- Sample Diluent (3 mL × 1 bottle)
- Sealing Films (2 pieces per 48-well; 2 pieces per 96-well)
- Sealed Bags (1 per configuration)
**Storage Conditions**
All components should be stored at 2–8°C. The kit has a shelf life of 6 months from the date of manufacture.
**Sample Preparation Guidelines**
Proper sample handling is essential for reliable results.
- **Serum**: Allow blood to clot at room temperature for 10–20 minutes, then centrifuge at 2000–3000 rpm for 20 minutes.
- **Plasma**: Use EDTA or sodium citrate as anticoagulant. Mix well and centrifuge similarly.
- **Urine**: Centrifuge at 2000–3000 rpm for 20 minutes.
- **Cell Culture Supernatant**: Centrifuge to remove debris before testing. For intracellular components, lyse cells by freezing and thawing.
- **Tissue Samples**: Homogenize in PBS, centrifuge, and collect supernatant. Store at 2–8°C after thawing.
**Important Notes**
- Avoid repeated freeze-thaw cycles.
- Do not use samples containing NaN3, as it may inhibit HRP activity.
- Ensure all reagents are equilibrated to room temperature before use.
- Always include blank wells and perform a standard curve.
- Use a pipette with accuracy and avoid cross-contamination.
- Handle all waste as biohazardous material.
**Procedure Summary**
1. Prepare standard dilutions and load them into the plate.
2. Add sample diluent and test samples to respective wells.
3. Incubate at 37°C for 30 minutes.
4. Wash the plate five times with diluted wash buffer.
5. Add enzyme-labeled reagent and incubate again.
6. Add TMB substrate and incubate for 15 minutes.
7. Stop the reaction and measure OD at 450 nm.
8. Calculate concentrations using a standard curve.
**Data Analysis**
Plot the standard curve using concentration vs. OD values. Determine the unknown sample concentration by interpolation or linear regression. Multiply by the dilution factor if applicable.
**Performance Characteristics**
- Intra-assay CV < 9%
- Inter-assay CV < 15%
- Detection range: 0.7 ng/mL – 22 ng/mL
- Correlation coefficient (R²) ≥ 0.92
**Safety and Handling**
Follow all laboratory safety protocols. Wear gloves and eye protection when handling reagents. Dispose of all waste properly.
This kit provides a reliable and efficient method for quantifying rat neutrophil elastase, making it ideal for research applications in immunology, inflammation studies, and disease biomarker analysis.
White Primer Moulded Door Moulded Panel doors
white primer molded panel door is an interior door that features a molded panel design and comes pre-primed with a white primer coat. These doors offer a combination of style, affordability, and ease of customization. Here are some key features and benefits of white primer molded panel doors:
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Molded panel design: White primer molded panel doors have a molded construction that creates raised or recessed panel designs on the surface. These panels add depth and visual interest to the door, mimicking the appearance of traditional solid wood doors.
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Pre-primed with white primer: The doors come with a white primer coat already applied to the surface. The primer serves as a base layer for paint, making it easier to achieve a smooth, finished look when painting the door in the desired color.
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Style versatility: White primer molded panel doors are available in various panel designs, such as 2-panel, 3-panel, 4-panel, or 6-panel configurations. This allows homeowners to choose a style that complements their interior decor and personal preferences.
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Affordability: Compared to solid wood doors, white primer molded panel doors are generally more affordable. They provide a cost-effective option for homeowners who desire the look of a traditional paneled door without the higher price tag.
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Easy customization: The white primer coat provides a blank canvas for customization. Homeowners can easily paint the door in any color or finish of their choice to match their interior design scheme. The primer layer helps the paint adhere properly and enhances the durability of the finish.
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Low maintenance: White primer molded panel doors typically require minimal maintenance. The painted surface is easy to clean with mild soap and water to keep it looking fresh.
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Consistency and stability: Molded panel doors are manufactured using engineered wood cores or composite materials that provide stability and reduce the likelihood of warping or twisting. This ensures that the doors maintain their shape and functionality over time.
It's important to note that white primer molded panel doors are intended for interior use only and are not suitable for exterior applications. They are commonly used for bedrooms, living rooms, closets, and other areas within the home.
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