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for Elisa Kit

Liquid Samples

• Serum: Collect in a sterile tube. Allow the blood to clot naturally for 120 minutes at room temperature or overnight at 2–8°C. Centrifuge at 3,000 rpm for 20 minutes at 2–8°C. Carefully collect the supernatant for immediate assay, or store at -20°C or -80°C for later use. Avoid repeated freeze-thaw cycles.
• Plasma: Choose EDTA or sodium heparin as the anticoagulant according to sample requirements. Mix thoroughly and centrifuge at 3,000 rpm for 20 minutes at 2–8°C within 30 minutes. Carefully collect the supernatant for immediate assay, or store at -20°C or -80°C for later use. Avoid repeated freeze-thaw cycles.
• Urine, Pleural/Abdominal Effusion, Cerebrospinal Fluid, and Saliva: Collect in a sterile tube and centrifuge at 3,000 rpm for 20 minutes at 2–8°C. Carefully collect the supernatant for immediate assay, or store at -20°C or -80°C. Avoid repeated freeze-thaw cycles.
• Cell Culture Supernatant: Collect in a sterile tube and centrifuge at 3,000 rpm for 20 minutes at 2–8°C. Carefully collect the supernatant for immediate assay, or store at -20°C or -80°C. Avoid repeated freeze-thaw cycles.

Solid Samples

• Animal Tissue Samples: Rinse tissue with pre-cooled PBS (0.01 M, pH = 7.4) to remove residual blood. Weigh and mince tissue into small pieces. Transfer the minced tissue into a glass homogenizer with an appropriate volume of PBS (typically a 1:9 w/v ratio, e.g., 1 g tissue to 9 mL PBS; adjust as needed and record the volume; adding protease inhibitors to the PBS is recommended). Grind thoroughly on ice/at low temperature, or use a tissue grinder. If further cell lysis is needed, perform sonication on the homogenate. Finally, centrifuge the homogenate at 5,000 × g for 10 minutes at 2–8°C and collect the supernatant for assay. For other tough tissues, grind thoroughly in liquid nitrogen if glass homogenizers or tissue grinders prove insufficient.
• Plant Tissue Samples: Rinse tissue with pre-cooled PBS (0.01 M, pH = 7.4) to remove residual soil and pesticides. Weigh and mince tissue into small pieces. Transfer the minced tissue into a glass homogenizer with an appropriate volume of PBS (typically a 1:9 w/v ratio, e.g., 1 g tissue to 9 mL PBS; adjust as needed and record the volume; adding protease inhibitors to the PBS is recommended). Grind thoroughly on ice/at low temperature, or use a tissue grinder. If further cell lysis is needed, perform sonication on the homogenate. Finally, centrifuge the homogenate at 5,000 × g for 10 minutes at 2–8°C and collect the supernatant. Aliquot the supernatant for single use, store at -20°C or -80°C, and avoid repeated freeze-thaw cycles.

Cell Samples

• Animal Cells: For adherent cells, wash gently with pre-cooled PBS and detach using trypsin. Collect cells by centrifugation at 1,000 × g for 5 minutes (suspension cells can be collected directly by centrifugation). Discard supernatant and wash cells 3 times with cold PBS. Resuspend cells in cold PBS at a density of 1 × 10^7 cells/mL. Disrupt cells thoroughly using a sonicator to release intracellular components. Centrifuge at 3,000 rpm for 20 minutes at 2–8°C, then carefully collect the supernatant for assay.
• Plant Cells: Dilute the cell suspension with PBS (pH 7.2–7.4) to reach approximately 1 × 10^6 cells/mL. Place on ice and disrupt cells thoroughly using a sonicator. Centrifuge at 3,000 rpm for 20 minutes at 2–8°C, and carefully collect the supernatant for assay.

Throat Swabs

Add 2 mL of PBS (pH 7.2–7.4) to dissolve the swab head and shake well. Remove the swab with forceps, squeezing out remaining liquid. Centrifuge at 2,000–3,000 rpm for approximately 20 minutes at 2–8°C, then carefully collect the supernatant. Keep one aliquot for testing and freeze the remainder for future use. If precipitation forms during storage, re-centrifuge prior to loading.

Sample Requirements

Perform extraction as soon as possible after sample collection, and run assays promptly following extraction. If assays cannot be run immediately, store samples at 2–8°C for use within 6 days. Otherwise, store at -20°C (≤1 month) or -80°C (≤2 months). Avoid repeated freeze-thaw cycles. If precipitation forms during storage, re-centrifuge prior to loading. Samples containing NaN3 cannot be analyzed, as NaN3 inhibits Horseradish Peroxidase (HRP) activity.