**Rabbit Tumor Necrosis Factor-α (TNF-α) ELISA Kit – Instructions for Use**
This reagent is intended for research purposes only. The kit is designed to quantitatively measure tumor necrosis factor alpha (TNF-α) in rabbit serum, plasma, and other biological fluids.
**Principle of the Assay**
The assay is based on the double-antibody sandwich ELISA method. A microplate is pre-coated with a specific antibody against rabbit TNF-α. After adding the sample, TNF-α binds to the immobilized antibody. An HRP-conjugated secondary antibody is then added, forming a complex of antibody-antigen-enzyme-labeled antibody. Following washing steps, the substrate TMB is introduced. Under the catalytic action of HRP, TMB turns blue and then changes to yellow upon acid addition. The intensity of the color is directly proportional to the concentration of TNF-α in the sample. Absorbance is measured at 450 nm using a microplate reader, and the TNF-α concentration is determined by comparing the results to a standard curve.
**Kit Components**
- Microtiter Plate: 1 × 48 or 1 × 96 wells
- Standard: 22.5 ng/L, 0.5 mL × 1 bottle
- Standard Diluent: 1.5 mL × 1 bottle
- Enzyme Label: 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
- Wash Buffer (20×): 20 mL × 20 times or 20 mL × 30 times
- Sealing Film: 2 pieces (for 48-well), 2 pieces (for 96-well)
- Storage Bag: 1 piece
**Storage Conditions**
All components should be stored at 2–8°C. Do not freeze unless specified.
**Sample Preparation and Handling**
1. **Serum**: Allow blood to clot at room temperature for 10–20 minutes, then centrifuge at 2000–3000 rpm for 20 minutes. Collect supernatant; if precipitate forms, re-centrifuge.
2. **Plasma**: Use EDTA or sodium citrate as anticoagulant. Mix for 10–20 minutes, then centrifuge at 2000–3000 rpm for 20 minutes. Collect supernatant.
3. **Urine**: Collect in a sterile tube and centrifuge at 2000–3000 rpm for 20 minutes. Carefully collect supernatant.
4. **Cell Culture Supernatant**: Centrifuge at 2000–3000 rpm for 20 minutes. For intracellular components, lyse cells via repeated freeze-thaw cycles, then centrifuge again.
5. **Tissue Samples**: Weigh the tissue, add PBS (pH 7.4), homogenize, and centrifuge at 2000–3000 rpm for 20 minutes. Collect supernatant. Store remaining samples at -20°C.
6. **General**: Process samples immediately after collection. If not tested right away, store at -20°C. Avoid repeated freezing and thawing.
7. **Note**: Samples containing NaN3 cannot be used, as it inhibits HRP activity.
**Important Notes**
Always follow the manufacturer’s instructions carefully. Ensure all reagents are brought to room temperature before use. Perform the assay in a clean environment to avoid contamination. Properly discard all waste materials according to local regulations.
Salt And Pepper Shaker
We are manufacturer and supplier of Stainless Steel Salt And Pepper Dispenser,Pepper Salt Shaker,Seasoning Jars,Stainless Steel Pepper Shaker,Spice Jars. And we located in Jiangmen, Guangdong, China. If any interested, please contact us for free.
Salt And Pepper Dispenser,Pepper Salt Shaker,Seasoning Jars,Stainless Steel Pepper Shaker,Spice Jars
Jiangmen Xinhui Zhancha Metal Products Co,. Ltd. , https://www.zchardware.com