**Human Mycoplasma pneumoniae (MP) ELISA Kit – For the Quantitative In Vitro Determination of Human MP Concentrations in Serum, Plasma, Cerebrospinal Fluid, Tissue Homogenate, and Other Body Fluids** *For Laboratory Research Use Only. Not Intended for Diagnostic or Therapeutic Procedures.* Before using this kit, please read the entire package insert carefully. This ELISA kit is designed for research purposes only and is not suitable for clinical diagnostic use. --- ### **Intended Use and Test Principle** The Human Mycoplasma pneumoniae (MP) ELISA Kit is intended for laboratory research use only. It is not approved for diagnostic or therapeutic applications. The assay is based on the enzyme-linked immunosorbent assay (ELISA) principle, which quantitatively measures MP concentrations in biological samples. The color change from blue to yellow is induced by the Stop Solution. A standard curve is generated by measuring the optical density (OD) of known MP concentrations, allowing the determination of MP levels in unknown samples by comparing their OD values to the standard curve. --- ### **Sample Collection and Storage** - **Serum**: Collect using 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 Biological Fluids**: Remove particulates by centrifugation. Assay immediately or store at -20°C. Ensure proper centrifugation and avoid hemolysis or contamination. --- ### **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** | 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 | *Note: Standard concentrations are 16, 8, 4, 2, 1, 0.5 U/L. If sample values exceed the highest standard, dilute with Sample Diluent and repeat the test. Do not substitute reagents between different kit lots. All components are matched for optimal performance.* --- ### **General Handling and Safety Instructions** - Allow all reagents and materials to reach room temperature (20–25°C) before use. - Do not thaw samples or reagents using water baths. - Do not use any reagent beyond its expiration date. - Always use deionized or distilled water for dilutions. - Keep microtiter plates in sealed bags until needed. Unused strips should be stored at 2–8°C with desiccant. - Use fresh pipette tips for each transfer to prevent cross-contamination. - Handle all blood-derived samples with caution, as they may carry infectious agents. - Dispose of waste properly—add sodium hypochlorite to liquid waste to a final concentration of 1.0% and allow it to stand for 30 minutes before disposal. - Substrate solution must be used before it turns bluish. - Chromogen B contains 20% acetone; keep away from heat or flame. --- ### **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. Add standards and samples in duplicate to the microtiter plate. 2. Add 50 μl of standard or sample to each well (blank well receives no addition). 3. Add 100 μl of HRP-conjugate reagent to all wells except the blank. 4. Wash the plate four times manually or automatically. - Manual: Aspirate contents, cover with adhesive strip, incubate for 60 minutes at 37°C. - Automated: Aspirate and wash four times using 1X Wash Buffer. 5. After washing, 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. 6. Add 50 μl of Stop Solution to each well. Read OD at 450 nm within 15 minutes. --- ### **Calculation and Interpretation** - Plot average OD values (450 nm) against standard concentrations to create a standard curve. - Subtract blank OD values before interpreting results. - Use graph paper or software to construct the curve. - Intersect the sample OD with the curve to determine MP concentration. - Variations in technique, timing, or reagent age can affect results. Each user should generate their own standard curve. --- ### **Performance Characteristics** - **Sensitivity:** <0.1 U/L - **Dynamic Range:** 0.5 U/L – 16 U/L - **Cross-Reactivity:** No significant cross-reactivity observed. - **Storage:** 2–8°C (frequent use); 6 months at -20°C. --- ### **Important Notes** - Always follow good laboratory practices when handling biological samples. - Never reuse reagents from different kit lots. - Maintain accurate records of all procedures and results. - This kit is not intended for human diagnostic use. *Please refer to the complete user manual for detailed instructions and safety information.*

Round Hot Pack(Bags)

Small hot compress bag,Heat packs reusable,Reuse heat packs

Ningbo Hejia Ice pack co. LTD, , https://www.cooling-pack.com