Analysis of the latest elisa kit - Database & Sql Blog Articles

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Component Analysis of the Latest ELISA Kit

The ELISA kit is a solid-phase sandwich enzyme-linked immunosorbent assay (ELISA), designed for high sensitivity and specificity in detecting target molecules. The process involves incubation, washing to remove unbound enzyme conjugates, followed by the addition of substrates A and B. The enzyme conjugate then catalyzes the reaction, producing a color change that can be measured spectrophotometrically.

Sample Preparation Guidelines

  1. Serum: Avoid any cell stimulation during sample collection. Use pyrogen-free and endotoxin-free tubes. After blood collection, separate red blood cells via centrifugation at 1000 x g for 10 minutes.
  2. Plasma: EDTA, citrate, or heparin plasma can be used. Centrifuge at 1000 x g for 30 minutes to remove the pellet.
  3. Cell Supernatant: Centrifuge at 1000 x g for 10 minutes to remove particles and polymer substances.
  4. Tissue Homogenate: Add tissue to an appropriate volume of physiological saline, chop it, and centrifuge at 1000 x g for 10 minutes to collect the supernatant.
  5. Storage: If not used immediately, store samples in small aliquots at -70°C to prevent repeated freezing. Avoid using hemolyzed or hyperlipidemic samples.

Kit Components

  • Pre-coated Plate: Anti-mouse interleukin-6 rabbit IgG, affinity-purified, 96-well plate.
  • Enzyme-labeled Antibody: HRP-conjugated anti-mouse IL-6 rabbit IgG, 30x concentrated, 0.4mL x 1.
  • Standard: Recombinant Mouse Interleukin-6, 0.5mL x 2.
  • EIA Buffer: Contains 1% BSA, 0.05% Tween 20, 30mL x 1.
  • Labeled Antibody Diluent: Contains 1% BSA, 0.05% Tween 20, 12mL x 1.
  • Reagent: TMB substrate, 15mL x 1.
  • Stop Solution: 1N sulfuric acid, 12mL x 1.
  • Concentrated Washing Solution: 40x concentrated, with 1% BSA, 0.05% Tween 20, 50mL x 1.

Equipment Required (Not Included in the Kit)

  • Microplate reader (450nm)
  • Micropipette and tips
  • Cylinder and beaker
  • Deionized water
  • Refrigerator (4°C)
  • Log/log coordinate paper
  • Absorbent paper
  • Test tubes (for standard dilution)
  • Incubator (37°C ± 1°C)
  • Wash bottle
  • Disposable reagent tubes (for concentrated enzyme-labeled antibody and color developer)

This detailed guide ensures accurate and consistent results when using the ELISA kit. Proper preparation and handling of samples are essential for reliable detection of interleukin-6 levels in various biological matrices.

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