**Rat Interleukin-2 (IL-2) Enzyme-Linked Immunosorbent Assay (ELISA) Kit – User Manual**
This reagent is intended solely for research purposes. The kit is designed to quantitatively determine the concentration of interleukin-2 (IL-2) in rat serum, plasma, cell culture supernatants, and other related liquid samples.
**Principle of the Assay**
The IL-2 ELISA kit employs the sandwich immunoassay technique. A microtiter plate is pre-coated with a specific monoclonal antibody against rat IL-2. After incubation with the sample, IL-2 present in the sample binds to the immobilized antibody. A biotinylated detection antibody is then added, followed by horseradish peroxidase (HRP)-conjugated streptavidin. The enzyme-substrate reaction is initiated using TMB (3,3',5,5'-tetramethylbenzidine), which changes color from clear to blue under HRP catalysis. The addition of a stop solution converts the color to yellow. The intensity of the yellow color is directly proportional to the amount of IL-2 in the sample. The absorbance is measured at 450 nm using a microplate reader, and the IL-2 concentration is determined by comparing the sample OD values to a standard curve.
**Kit Components**
- 48-well configuration:
- Microtiter plate (1 × 48 wells)
- Standard (1800 ng/L, 0.5 mL × 1 bottle)
- Standard diluent (1.5 mL × 1 bottle)
- Enzyme-labeled antibody (3 mL × 1 bottle)
- Sample diluent (3 mL × 1 bottle)
- TMB substrate (3 mL × 1 bottle)
- Stop solution (3 mL × 1 bottle)
- Wash buffer (20 mL × 20 times, 1 bottle)
- Plate sealer (2 pieces)
- Sealing film (1 piece)
- 96-well configuration:
- Microtiter plate (1 × 96 wells)
- Standard (1800 ng/L, 0.5 mL × 1 bottle)
- Standard diluent (1.5 mL × 1 bottle)
- Enzyme-labeled antibody (6 mL × 1 bottle)
- Sample diluent (6 mL × 1 bottle)
- TMB substrate (6 mL × 1 bottle)
- Stop solution (6 mL × 1 bottle)
- Wash buffer (20 mL × 30 times, 1 bottle)
- Plate sealer (2 pieces)
- Sealing film (1 piece)
**Storage Instructions**
All components should be stored at 2–8°C. The kit is stable for up to 12 months when stored properly. Avoid freezing and protect from light.
**Notes**
It is recommended to prepare fresh dilutions of standards and samples before use. Ensure all reagents are brought to room temperature before use. Proper pipetting techniques and thorough washing steps are essential for accurate results. Always follow the manufacturer’s instructions and safety guidelines when handling chemicals.
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