Human LTA ELISA Kit

**Human LTA ELISA Kit – For the quantitative in vitro determination of Human Lung Tumor Antigen concentrations in serum, plasma, celiac 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 ELISA (Enzyme-Linked Immunosorbent Assay) is intended for laboratory research purposes only and is not suitable for diagnostic or therapeutic use. The LTA ELISA Kit allows for the accurate quantification of Human Lung Tumor Antigen (LTA) in various biological samples. The principle of the assay involves the use of a standard curve generated from known concentrations of LTA. By comparing the optical density (OD) readings of the sample to the standard curve, the concentration of LTA in the sample can be precisely determined. **INTENDED USE AND TEST PRINCIPLE** This LTA ELISA Kit is designed for research use only. The Stop Solution changes the reaction color from blue to yellow, and the intensity of the color is proportional to the amount of LTA present in the sample. Calibration standards are included in the kit to create a reference curve for accurate quantification. **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. Analyze immediately or 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 repeated freeze-thaw cycles. - **Cell culture supernatants, tissue homogenates, and other biological fluids**: Remove particulates by centrifugation. Analyze immediately or store at -20°C. Ensure no hemolysis or granules are present. **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** All reagents should be stored at 2–8°C. Check the expiration date on the label before use. - MicroELISA Strip Plate: 12×8 strips / 12×4 strips - Standard (6 vials): 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 units - User Manual: 1 unit - Sealed Bags: 1 unit **Note:** Standard concentrations: 4000, 2000, 1000, 500, 250, 125 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 kits. All components are calibrated for optimal performance. 2. Allow all reagents and materials to reach room temperature (20–25°C) before use. Do not use water baths for thawing. 3. Do not use reagents past their expiration date. 4. Only use deionized or distilled water for dilutions. 5. Keep microtiter plates in sealed bags until needed. Unused strips should be stored with desiccant at 2–8°C. 6. Use fresh pipette tips for each transfer to avoid cross-contamination. 7. Handle all blood-derived samples with care, as they may carry infectious agents. Follow proper biosafety protocols. 8. Dispose of all samples and waste according to local regulations. 9. Waste should be treated with sodium hypochlorite to a final concentration of 1.0% and left for at least 30 minutes to inactivate viruses. 10. Substrate solutions are sensitive to contamination. Store away from heat and flame. 11. Allow all reagents to warm to room temperature before use. **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 the assay. 2. Add 50 µl of standard or sample to appropriate wells (excluding blank). Cover with adhesive strip and incubate for 60 minutes at 37°C. 3. Wash the plate 4 times using either manual or automated washing methods. - **Manual Washing**: Aspirate contents, fill with 1X Wash Solution, and aspirate again. Repeat four times. Invert and blot dry. - **Automated Washing**: Aspirate and wash four times. Adjust brush settings and fill volume to 350 µL per well. 4. Add 50 µl of Chromogen A and 50 µl of Chromogen B to each well. Mix gently and incubate for 15 minutes at 37°C, protecting from light. 5. Add 50 µl of Stop Solution to each well. Read OD at 450 nm using a microplate reader. 6. Plot the average OD values of the standards against their concentrations to generate a standard curve. 7. Calculate the mean OD for each sample and subtract the blank value. Use the standard curve to determine the LTA concentration in each sample. **PERFORMANCE DATA** - Intra-assay CV (%) and Inter-assay CV (%) < 15%. - Assay range: 125 pg/ml – 4000 pg/ml. - Sensitivity: <100 pg/ml. - No significant cross-reactivity or interference observed. **STORAGE** - Store at 2–8°C for frequent use; up to six months at -20°C. **NOTES** Always follow safety guidelines and proper lab procedures when handling biological samples and reagents. Each user should generate their own standard curve for best results. For any questions or technical support, refer to the manufacturer’s instructions or contact their customer service team.

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