**Rat Interleukin 2 (IL-2) Enzyme-Linked Immunosorbent Assay (ELISA) Kit – Instruction Manual**
This reagent is intended for research purposes only. It is designed to quantify the levels of interleukin-2 (IL-2) in rat serum, plasma, cell culture supernatants, and other related biological fluids.
**Experimental Principle**
The kit utilizes a double-antibody sandwich ELISA method to detect IL-2 in the sample. A microplate is pre-coated with a purified monoclonal antibody specific to rat IL-2. The sample is added to the wells, allowing IL-2 to bind to the immobilized antibody. After washing, a horseradish peroxidase (HRP)-labeled anti-IL-2 antibody is introduced, forming an antibody-antigen-enzyme-labeled antibody complex. Following a thorough wash step, the TMB substrate is added, which changes color under the catalytic action of HRP. The reaction is stopped with an acidic solution, resulting in a yellow color. The intensity of the color is directly proportional to the concentration of IL-2 in the sample. The optical density (OD) is measured at 450 nm using a microplate reader, and the IL-2 concentration is determined by comparing the OD values to a standard curve.
**Kit Components**
- **Coated Plate**: 1 × 48 or 1 × 96 wells
- **Standard**: 0.5 ml × 1 bottle (1800 ng/L)
- **Standard Diluent**: 1.5 ml × 1 bottle
- **Enzyme Labeled Antibody**: 3 ml × 1 bottle
- **Sample Diluent**: 3 ml × 1 bottle
- **TMB Substrate (A & B)**: 3 ml × 1 bottle each
- **Stop Solution**: 3 ml × 1 bottle
- **Washing Buffer (Concentrated)**: 20 ml × 20 times or 20 ml × 30 times
- **Sealing Film**: 2 pieces (for 48-well), 2 pieces (for 96-well)
- **Sealed Bag**: 1 piece
**Storage Instructions**
All components should be stored at 2–8°C. Ensure the kit is kept away from light and moisture. Do not freeze unless specified.
**Notes for Use**
Before use, bring all reagents to room temperature. Follow the protocol carefully to ensure accurate results. Proper calibration with standards is essential for reliable quantification. Always perform the assay in duplicate or triplicate to minimize variability.
**Important**
This kit is not intended for diagnostic use. The results obtained are for research purposes only.
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