Rat neutrophil elastase (NE) ELISA kit instruction manual

**Rat Neutrophil Elastase (NE) ELISA Kit – Instructions for Use** This kit is intended for research purposes only and is designed to quantitatively measure Rat Neutrophil Elastase (NE) in serum, plasma, and other biological fluids. The method used is a double-antibody sandwich ELISA, which allows for accurate detection of NE levels. **Principle of the Assay** The assay involves coating a microtiter plate with a specific antibody against Rat Neutrophil Elastase. After incubation with the sample, a second HRP-conjugated antibody binds to the captured NE, forming an antibody-antigen-enzyme complex. Following washing steps, the substrate TMB is added, producing a color change that correlates with the amount of NE present. 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 concentration of NE in the samples. **Kit Components** The kit includes all necessary reagents for performing the assay. It comes in 48-well or 96-well configurations, with components such as: - Coated plates - Standard solutions (with different concentrations) - Enzyme-labeled antibodies - Substrates (TMB solution A and B) - Washing buffer - Sample diluent - Stop solution All reagents should be stored at 2–8°C and used within the expiration date of 6 months from the date of manufacture. **Sample Preparation Guidelines** Proper handling of samples is essential for accurate results. 1. **Serum**: Allow blood to clot at room temperature for 10–20 minutes, then centrifuge at 2000–3000 rpm for 20 minutes. Collect the supernatant carefully. 2. **Plasma**: Use EDTA or sodium citrate as anticoagulant, mix well, and centrifuge similarly. 3. **Urine**: Centrifuge for 20 minutes, collect the supernatant. 4. **Cell culture supernatant**: Centrifuge after collection; for intracellular components, lyse cells by repeated freezing and thawing before centrifugation. 5. **Tissue homogenate**: Homogenize tissue in PBS, centrifuge, and collect the supernatant. 6. Avoid samples containing NaN3, as it may interfere with the HRP enzyme activity. **Procedure Summary** 1. Prepare standard dilutions and load them into designated wells. 2. Add sample diluent and test samples to respective wells. 3. Incubate at 37°C for 30 minutes. 4. Wash the plate thoroughly. 5. Add the enzyme-labeled antibody and incubate again. 6. Add TMB substrate and incubate for 15 minutes in the dark. 7. Stop the reaction with stop solution. 8. Measure OD values at 450 nm. 9. Calculate NE concentration using a standard curve. **Notes and Recommendations** - Equilibrate the kit at room temperature for 15–30 minutes before use. - Ensure all reagents are properly mixed and handled. - Always include blank wells and perform duplicate measurements. - If sample concentration exceeds the standard range, dilute the sample accordingly. - Avoid cross-contamination by using a new sealing film for each experiment. - Keep all reagents away from light and handle with care. - Follow the manufacturer’s instructions strictly for reliable results. **Data Interpretation** Plot the standard curve using known concentrations and corresponding OD values. Determine the unknown sample concentration by interpolation or linear regression. Multiply the result by the dilution factor if applicable. **Performance Characteristics** - The correlation coefficient (R²) of the standard curve is ≥ 0.92. - Intra-batch and inter-batch variability is less than 9% and 15%, respectively. - Detection range: 0.7 ng/mL to 22 ng/mL. **Storage Conditions** Store the unopened kit at 2–8°C. Once opened, store unused components in sealed bags and follow the recommended storage conditions. The kit is valid for 6 months from the date of receipt.

Bathroom Cabinet

Bathroom cabinet is a piece of furniture designed specifically for use in bathrooms. It is primarily used for storage and organization of various bathroom essentials, such as toiletries, towels, and other personal care items. Bathroom cabinets come in a wide range of styles, sizes, and materials to suit different bathroom designs and storage needs.

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