**Chicken Whitefly Antibody (PD) ELISA Test Kit – User Manual**
This ELISA kit is designed for the quantitative detection of Chicken Whitefly Antibody (PD) in biological samples using a sandwich ELISA method. The principle involves the use of pre-coated microplates with specific capture antibodies, followed by sequential incubation with standards, samples, and HRP-conjugated detection antibodies. After washing, TMB substrate is added, and color development occurs under peroxidase activity, resulting in a blue to yellow color change. The intensity of the color is directly proportional to the concentration of PD antibody in the sample. Absorbance is measured at 450 nm using a microplate reader, allowing for accurate quantification.
**Sample Collection and Preparation**
1. **Serum**: Collect blood in pyrogen-free tubes. Centrifuge at 3000 rpm for 10 minutes to separate serum from red blood cells.
2. **Plasma**: Use anticoagulants such as EDTA, citrate, or heparin. Centrifuge at 3000 rpm for 30 minutes and collect the supernatant.
3. **Cell Supernatant**: Centrifuge at 3000 rpm for 10 minutes to remove debris.
4. **Tissue Homogenate**: Homogenize tissue in physiological saline, then centrifuge at 3000 rpm for 10 minutes to obtain the supernatant.
5. **Storage**: Store samples at -20°C if not tested immediately. Avoid repeated freeze-thaw cycles. Thaw at room temperature before use.
**Required Equipment**
- Microplate reader (450 nm)
- Precision pipettes (0.5–10 µL, 2–20 µL, 20–200 µL, 200–1000 µL)
- Incubator at 37°C
**Important Notes**
- Store the kit at 2–8°C. Allow it to equilibrate at room temperature for 20 minutes before use.
- If the wash buffer crystallizes after removal from the fridge, warm it gently in a water bath before use.
- Unused strips should be returned to the ziplock bag and stored at low temperature.
- No dilution is required for pre-treated samples; add 10 µL directly.
- Follow the protocol strictly regarding timing, volume, and sequence.
- Shake all reagents well before use.
**Kit Components**
| Component | 96-well | 48-well | Notes |
|----------|---------|---------|-------|
| Microporous Plate | 12 × 8 | 12 × 4 | - |
| Standard (200 pg/mL) | 0.5 mL | 0.5 mL | - |
| Diluted Standards | 6 mL | 3 mL | As per instructions |
| Sample Diluent | 6 mL | 3 mL | As per instructions |
| Detection Antibody-HRP | 6 mL | 3 mL | - |
| 20× Wash Buffer | 20 mL | 20 mL | Dilute with distilled water |
| Substrate A | 6 mL | 3 mL | - |
| Substrate B | 6 mL | 3 mL | - |
| Stop Solution | 6 mL | 3 mL | - |
| Seal Film | 2 sheets | 2 sheets | - |
| Instructions | 1 part | 1 part | - |
| Zip Lock Bag | 1 piece | - | - |
**Reagent Preparation**
- **20× Wash Buffer**: Dilute 1 part with 19 parts of distilled water.
**Washing Procedure**
- **Manual**: Fill each well with washing solution, let stand for 1 minute, discard, and repeat 5 times.
- **Automated**: Add 350 µL of wash solution, soak for 1 minute, and repeat 5 times.
**Assay Procedure**
1. Remove the required microtiter strips from the foil pouch after 20 minutes at room temperature. Store the rest in a sealed bag at 4°C.
2. Set up standard, sample, and blank wells.
3. Add 50 µL of standard solutions (ranging from 200 pg/mL to 0 pg/mL).
4. Add 10 µL of sample and 40 µL of diluent to each sample well.
5. Add 50 µL of HRP-labeled detection antibody to each well. Seal and incubate at 37°C for 60 minutes.
6. Wash 5 times with washing solution.
7. Add 50 µL of TMB substrate A and B, incubate in the dark for 15 minutes.
8. Add 50 µL of stop solution and measure OD at 450 nm within 15 minutes.
**Data Analysis**
Plot standard concentrations against their corresponding OD values in Excel. Generate a standard curve and calculate sample concentrations based on the regression equation.
**Kit Performance**
- **Accuracy**: R² ≥ 0.9900
- **Sensitivity**: <1.0 pg/mL
- **Specificity**: No cross-reactivity with other analogs
- **Reproducibility**: CV <15% between plates
- **Storage**: 2–8°C, protected from light and moisture
- **Shelf Life**: 6 months
- **Detection Range**: 6.25 pg/mL – 200 pg/mL
**Disclaimer**
This kit is intended for research purposes only. Not suitable for clinical trials or animal testing. The company is not responsible for any misuse or deviation from the instructions. Always follow the protocol carefully. Do not mix reagents from different batches. Any errors due to improper handling are the responsibility of the user.
A gas grill is a type of outdoor grill that uses propane or natural gas as a fuel source to cook food. It typically has a metal cooking grate, burners, and a hood or lid to trap heat and smoke. Gas grills are popular because they are easy to use and offer precise temperature control, making them ideal for grilling a variety of foods, from burgers and hot dogs to vegetables and seafood. They can also be used year-round, as they do not require charcoal or wood for fuel. Some gas grills also have additional features such as side burners, rotisserie attachments, and built-in thermometers.
Gas Grill ,Portable Propane Grill,Char Broil Bbq,Small Propane Grill
NingBo AoYue Technology Co., Ltd. , https://www.aoyue-tech.com