**Bovine Parainfluenza Virus Type III Antigen (PIV-III-Ag) ELISA Kit Instruction Manual**
This reagent is intended for research purposes only and is not for use in diagnostic procedures. The kit is designed to detect the presence of Bovine Parainfluenza Virus Type III Antigen (PIV-III-Ag) in bovine serum, plasma, and related biological samples.
**Experimental Principle**
The assay is based on a double-antibody sandwich ELISA method. A microtiter plate pre-coated with a specific antibody against PIV-III-Ag is used as a solid phase. The sample is added, allowing any PIV-III-Ag present to bind to the immobilized antibody. After washing to remove unbound materials, a horseradish peroxidase (HRP)-labeled secondary antibody is added, forming an immune complex. Following another wash, the substrate TMB is introduced. In the presence of HRP, TMB turns blue, and the reaction is stopped by adding an acidic solution, which changes the color to yellow. The optical density (OD) at 450 nm is measured using a microplate reader. By comparing the OD value to the cutoff value, the presence or absence of PIV-III-Ag in the sample can be determined.
**Kit Components**
The kit is available in 48-well and 96-well configurations.
- **Coated Microplate**: 1 × 48 or 1 × 96 wells
- **Sealing Film**: 2 pieces (for 48-well), 2 pieces (for 96-well)
- **Sealed Bag**: 1 piece
- **Negative Control**: 0.5 ml × 1 bottle
- **Positive Control**: 0.5 ml × 1 bottle
- **Enzyme-labeled Antibody**: 3 ml × 1 bottle (48-well), 6 ml × 1 bottle (96-well)
- **Sample Diluent**: 3 ml × 1 bottle (48-well), 6 ml × 1 bottle (96-well)
- **Developer A**: 3 ml × 1 bottle (48-well), 6 ml × 1 bottle (96-well)
- **Developer B**: 3 ml × 1 bottle (48-well), 6 ml × 1 bottle (96-well)
- **Stop Solution**: 3 ml × 1 bottle (48-well), 6 ml × 1 bottle (96-well)
- **Washing Solution (Concentrated)**: 20 ml × 20 times (for 48-well), 20 ml × 30 times (for 96-well)
**Storage Instructions**
All components should be stored at 2–8°C. Avoid freezing. Ensure the kit is used within the expiration date stated on the label.
**Precautions**
- Do not mix reagents from different batches.
- Avoid contact with skin and mucous membranes.
- Use pipettes with filter tips to prevent cross-contamination.
- All operations should be performed in a clean and controlled environment.
This manual provides detailed instructions for accurate and reliable detection of PIV-III-Ag in bovine samples. Follow all steps carefully to ensure optimal performance of the ELISA kit.
DL-Valine
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