Component Analysis of the Latest ELISA Kit
The ELISA kit is a solid-phase sandwich enzyme-linked immunosorbent assay (ELISA). It involves incubation and washing steps to remove unbound enzyme conjugates, followed by the addition of substrates A and B. The enzyme conjugate then produces a color change, which can be measured to determine the concentration of the target analyte.
ELISA Kit Composition
- Serum: Avoid any cell stimulation during the process. Use pyrogen-free and endotoxin-free tubes. After collecting blood, separate red blood cells quickly and carefully by centrifuging at 1000 x g for 10 minutes.
- Plasma: EDTA, citrate, or heparin plasma can be used. Centrifuge at 1000 x g for 30 minutes to remove the pellet.
- Cell Supernatant: Centrifuge at 1000 x g for 10 minutes to remove particles and polymers.
- Tissue Homogenization: Add tissue to an appropriate amount of physiological saline, chop it, and centrifuge at 1000 x g for 10 minutes. Take the supernatant for testing.
- Storage: If not used immediately, divide the sample into small parts and store at -70°C to avoid repeated freezing. Try to avoid hemolyzed or hyperlipidemic samples.
ELISA Kit Components
- 1. Pre-coated plate: Anti-mouse interleukin-6 rabbit IgG, affinity purified, 96T
- 2. Enzyme-labeled antibody: HRP-labeled anti-mouse interleukin-6 rabbit IgG, affinity purified, 30 times concentrated, 0.4mL x 1
- 3. Standard: Recombinant Mouse Interleukin-6, 0.5mL x 2
- 4. EIA Buffer: Contains 1% BSA, 0.05% Tween 20, BPS, 30mL x 1
- 5. Labeled Antibody Diluent: Contains 1% BSA, 0.05% Tween 20, BPS, 12mL x 1
- 6. Reagent: TMB substrate, 15mL x 1
- 7. Stop solution: 1N sulfuric acid, 12mL x 1
- 8. Concentrated washing solution: 40 times concentrated, with 1% BSA, 0.05% Tween 20, BPS, 50mL x 1
Equipment Required for the Experiment (Not Included in the Kit)
- Microplate reader (450nm)
- Micropipette and tips
- Cylinder and beaker
- Deionized water
- Refrigerator (4°C)
- Coordinate paper (log/log)
- Absorbent paper
- Test tube (for standard dilution)
- Incubator (37°C ± 1°C)
- Wash bottle (for washing)
- Disposable reagent tubes (for concentrated enzyme-labeled antibody and color developer)
This comprehensive ELISA kit provides all necessary reagents and components for accurate and reliable detection of mouse interleukin-6. Proper handling and storage of samples are essential to ensure optimal results. Make sure all equipment is clean and calibrated before starting the experiment.
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