Human LTA ELISA Kit

**Human LTA ELISA Kit – For the Quantitative In Vitro Determination of Human Lung Tumor Antigen Concentrations in Serum, Plasma, Cerebrospinal Fluid, Tissue Homogenate, and Other Biological Fluids** **For Laboratory Research Use Only. Not for Diagnostic or Therapeutic Procedures.** Before using this product, please read the entire package insert carefully. This ELISA (Enzyme-Linked Immunosorbent Assay) is intended for research purposes only and is not suitable for diagnostic or clinical use. **Intended Use and Test Principle** This LTA ELISA Kit is designed to quantitatively measure Human Lung Tumor Antigen (LTA) concentrations in various biological samples. The test works by using a set of calibration standards that are run alongside the samples. A standard curve is generated by plotting optical density (OD) values against known LTA concentrations. The sample LTA concentration is then determined by comparing its OD value to the standard curve. **Sample Collection and Storage** - **Serum**: Collect using a serum separator tube. Allow clotting for 2 hours at room temperature or overnight at 4°C before centrifuging at 2000×g for 20 minutes. Assay immediately or aliquot and store at -20°C. Avoid repeated freeze-thaw cycles. - **Plasma**: Collect with heparin as an anticoagulant. Centrifuge within 30 minutes at 2000×g, 2–8°C. Store at -20°C. - **Cell Culture Supernatant, Tissue Homogenate, and Other Biological Fluids**: Remove particulates by centrifugation. Assay immediately or store at -20°C. Ensure no hemolysis or debris remains. **Materials Required but Not Supplied** 1. Incubator 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 (Stored at 2–8°C)** | Reagent | 96 Determinations | 48 Determinations | |---------|-------------------|-------------------| | Microtiter 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: 4000, 2000, 1000, 500, 250, 125 pg/ml. If sample values exceed the highest standard, dilute with Sample Diluent and retest.* **Precautions** 1. Do not mix reagents from different kit lots. Each component is 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 past their expiration date. 4. Use only deionized or distilled water for dilution. 5. Keep microtiter plates in their sealed bags until ready for use. Unused strips should be stored with desiccant at 2–8°C. 6. Use fresh pipette tips for each transfer to prevent cross-contamination. 7. All blood-derived products should be treated as potentially infectious. Follow strict lab safety protocols. 8. Dispose of all samples and waste properly. Waste should be treated with sodium hypochlorite (final concentration 1.0%) for at least 30 minutes before disposal. 9. Substrate solutions may be easily contaminated. Avoid exposure to heat or flame. 10. Always allow reagents to reach 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 1 month. **Assay Procedure** 1. Prepare all reagents before starting. Add 50 µl of standard or sample to appropriate wells (excluding blank). Cover with adhesive strip and incubate for 60 minutes at 37°C. 2. Wash the microtiter plate 4 times (manual or automated). After final wash, invert and blot dry. 3. 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. 4. Add 50 µl of Stop Solution to each well. Read OD at 450 nm using a microplate reader. 5. Generate a standard curve by plotting average OD values (450 nm) against standard concentrations. Determine sample LTA levels by comparing their OD to the curve. 6. Ensure all OD values are corrected by subtracting the blank mean. 7. Intra-assay and inter-assay CV% are <15%. 8. Assay range: 125 pg/ml – 4000 pg/ml. 9. Sensitivity: <100 pg/ml. 10. No significant cross-reactivity or interference observed. **Storage** - Store at 2–8°C for frequent use; up to 6 months at -20°C. **Important Note** This kit is intended for research use only. It should not be used in diagnostic procedures. Always follow proper lab safety and disposal guidelines.

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