Type of experiment: Sandwich method detection range: 2.5 pg/mL – 80 pg/mL
Minimum detection limit: 0.1 pg/mL
Applications: Serum, plasma, tissue homogenate, cell culture supernatant or other related fluids (this product is for laboratory research and non-clinical use only)
Experimental principle
The kit uses a double antibody one-step sandwich enzyme-linked immunosorbent assay (ELISA). To the coated microwells pre-coated with canine tumor necrosis factor alpha (TNF-α) capture antibody, specimens, standards, and HRP-labeled detection antibodies were sequentially added, incubated and thoroughly washed. Using the substrate TMB to develop color, TMB is converted to blue under the catalysis of peroxidase and converted to the final yellow color by the action of an acid. The color depth is positively correlated with canine tumor necrosis factor alpha (TNF-α) in the sample. The absorbance (OD value) was measured at 450 nm using a microplate reader to calculate the sample concentration.
Precautions
1. Incubate strictly in accordance with the specified time and temperature to ensure accurate results. All reagents must reach room temperature 20-25 °C before use. Store the reagents refrigerated immediately after use.
2. Incorrect cleaning can result in inaccurate results. Make sure to drain the liquid in the well as much as possible before adding the substrate. Do not let the micropores dry during the incubation.
3. Eliminate residual liquid and fingerprints on the bottom of the board, otherwise it will affect the OD value.
4. The substrate coloring solution should be colorless or very light, and the substrate liquid that has turned blue cannot be used.
5. Avoid cross-contamination of reagents and specimens to avoid erroneous results.
6. Avoid direct exposure to strong light during storage and incubation.
7. After balancing to room temperature, open the sealed bag to prevent the water droplets from condensing on the cold slats.
8. Any reagents should not be exposed to strong gases emitted by bleaching solvents or bleaching solvents. Any bleaching component will destroy the biological activity of the reagents in the kit.
9. Cannot use expired products.
10. If the disease is likely to spread, all samples should be managed and the sample and test device processed in accordance with the prescribed procedures.
Experimental result calculation
The OD value of the tested standard is the abscissa, the concentration value of the standard is the ordinate, the standard curve is drawn on the coordinate paper or with the relevant software, and the linear regression equation is obtained, and the OD value of the sample is substituted into the equation to calculate the sample. concentration.
(This picture is for reference only)
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