The RayBio^® Mouse IL-6 ELISA kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of mouse Interleukin 6 in serum, plasma and cell culture supernatants. This assay employs an antibody specific for mouse coated on a 96-well plate. Standards and samples are pipetted into the wells and IL-6 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-Mouse IL-6 antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of IL-6 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm

Interleukin 6 is a pleiotropic cytokine that regulates cell growth and differentiation of various tissues, and plays an important role in the immune response and acute phase reactions. It acts as both a pro-inflammatory and anti-inflammatory cytokine. It is secreted by T cells and macrophages to stimulate an immune response, e.g., during infection and after trauma, especially burns or other tissue damage leading to inflammation. IL-6's role as an anti-inflammatory cytokine is mediated through its inhibitory effects on TNF-α and IL-1, and activation of IL-1Rα and IL-10.

• Strip plates and additional reagents allow for use in multiple experiments
• Pre-coated microplates for better reproducibility and efficiency
• Fully optimized for serum, plasma, cell culture medium, and lysates
• Quantitative protein detection
• Establishes normal range
• The best products for confirmation of multiplex array data

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Stop Solution
• Assay Diluent(s)
• Lyophilized Standard
• Biotinylated Detection Antibody
• Streptavidin-Conjugated HRP
• TMB One-Step Substrate

• Distilled or deionized water
• Precision pipettes to deliver 2 µL to 1 µL volumes
• Adjustable 1-25 µL pipettes for reagent preparation
• 100 µL and 1 liter graduated cylinders
• Tubes to prepare standard and sample dilutions
• Absorbent paper
• Microplate reader capable of measuring absorbance at 450nm
• Log-log graph paper or computer and software for ELISA data analysis

1. Prepare all reagents, samples and standards as instructed in the manual.
2. Add 100 µL of standard or sample to each well.
3. Incubate 2.5 h at RT or O/N at 4°C.
4. Add 100 µL of prepared biotin antibody to each well.
5. Incubate 1 h at RT.
6. Add 100 µL of prepared Streptavidin solution to each well.
7. Incubate 45 min at RT.
8. Add 100 µL of TMB One-Step Substrate Reagent to each well.
9. Incubate 30 min at RT.
10. Add 50 µL of Stop Solution to each well.
11. Read at 450 nm immediately.