Mouse Tumor Necrosis Factor β (TNF-β) ELISA Test Kit This reagent is for research use only
Test principle: The TNF-β kit is a solid-phase sandwich method enzyme-linked immunosorbent assay (ELISA). Standards with known TNF-β concentration and samples with unknown concentrations are added to the microplate for detection. Incubate TNF-β and biotin-labeled antibody first. After washing, HRP labeled with avidin was added. After incubation and washing, the unbound enzyme conjugate is removed, and then substrates A and B are added to act simultaneously with the enzyme conjugate. Produce colors. The color depth is proportional to the concentration of TNF-β in the sample.
Bring your own materials:
1. Distilled water.
2. Sampler: 5ul, 10ul, 50ul, 100ul, 200ul, 500ul, 1000ul.
3. Oscillator and magnetic stirrer, etc.
4. Sample collection, processing and storage methods: Serum ... Avoid any cell stimulation during the operation. Use test tubes free of pyrogens and endotoxins.
After collecting blood, centrifuge at 1000 × g for 10 minutes to quickly and carefully separate the red blood cells.
2. Plasma ... EDTA, citrate and heparin plasma can be used for detection. Centrifuge at 1000 × g for 30 minutes to remove particles.
3. Cell supernatant ... centrifuge at 1000 × g for 10 minutes to remove particles and polymer.
4. Tissue homogenate ... Mash the tissue with appropriate amount of saline. Centrifuge at 1000 × g for 10 minutes, and take the supernatant.
5. Storage ... If the sample is not used immediately, it should be divided into small parts -70 ℃ to avoid repeated freezing. If possible, do not use hemolysis or hyperlipidemia. If there are a large number of particles in the serum, centrifuge or filter before testing. Do not thaw at 37 ° C or higher. Thaw at room temperature and ensure that the sample is thawed evenly and adequately.
Operation notes:
â— Reagents should be stored according to the label instructions and returned to room temperature before use. The sparse standards should be discarded and cannot be stored.
â— Slats not used in the experiment should be immediately returned to the packaging bag, sealed and stored to avoid deterioration.
â— Any unused reagents should be packed or covered. Do not mix reagents of different batches. Use before warranty.
â— Use a disposable pipette tip to avoid cross-contamination, and avoid using a sampler with a metal part when drawing the stop solution and the substrates A and B.
â— Use a clean plastic container to prepare the washing solution. Mix all components and samples in the kit thoroughly before use.
â— Substrate A should evaporate to avoid opening the lid for a long time. Substrate B is sensitive to light and avoid prolonged exposure to light. Avoid contact with hands, it is toxic. The OD value should be read immediately after the experiment is completed.
â— The order of adding reagents should be consistent to ensure the same incubation time of all reaction wells.
â— Perform the incubation operation in accordance with the time, amount and sequence of the liquid indicated in the manual.
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