Rat tumor necrosis factor beta (TNF-β) Eelisa kit instruction manual

**Rat Tumor Necrosis Factor Beta (TNF-β) ELISA Kit – User Manual** **Kit Specifications:** This ELISA kit is available in 48-well or 96-well configurations. It includes: - Standard dilution: 1.5 ml × 1 vial - Enzyme standard reagent: 3 ml × 1 vial (for 48 wells) or 6 ml × 1 vial (for 96 wells) - Sealing film: 2 pieces (for 48 wells) or 2 pieces (for 96 wells) - Standard: 0.5 ml × 1 vial, concentration: 2700 ng/L - Sample diluent: 3 ml × 1 vial (48) / 6 ml × 1 vial (96) - Developer A: 3 ml × 1 vial (48) / 6 ml × 1 vial (96) - Chromogen B: 3 ml × 1 vial (48) / 6 ml × 1 vial (96) - Stop solution: 3 ml × 1 vial (48) / 6 ml × 1 vial (96) - Concentrated washing solution: 20 ml × 20 times (48) or 20 ml × 30 times (96) **Storage Conditions and Shelf Life:** - Store the kit at 2–8°C - Shelf life: 6 months from the date of manufacture **Purpose:** This ELISA kit is designed for the quantitative determination of tumor necrosis factor beta (TNF-β) in biological samples such as serum, plasma, urine, cell culture supernatants, and tissue homogenates. **Principle of Operation:** The kit employs a sandwich ELISA method. The microtiter plate is pre-coated with anti-TNF-β antibodies. After adding the sample, TNF-β binds to the immobilized antibody. An HRP-conjugated secondary antibody is then added, forming an immune complex. After washing, TMB substrate is added, leading to a color change that is proportional to the amount of TNF-β present. The reaction is stopped, and the absorbance is measured at 450 nm. A standard curve is plotted using known concentrations, and the sample concentration is calculated accordingly. **Sample Preparation Instructions:** 1. **Serum:** Allow blood to clot at room temperature, centrifuge at 2000–3000 rpm for 10–20 minutes, and collect the supernatant. 2. **Plasma:** Use EDTA or sodium citrate as anticoagulant. Mix well, centrifuge, and collect the supernatant. 3. **Urine:** Centrifuge at 2000–3000 rpm for 10–20 minutes, and collect the supernatant. 4. **Cell Culture Supernatant:** Centrifuge at 2000–3000 rpm for 10–20 minutes. For intracellular components, lyse cells by repeated freeze-thaw cycles, then centrifuge again. 5. **Tissue Homogenate:** Weigh the tissue, add PBS, homogenize, centrifuge, and collect the supernatant. 6. **General Notes:** Samples should be processed as soon as possible after collection. If not tested immediately, store at -20°C, avoiding repeated freezing and thawing. Do not use samples containing NaN3, as it inhibits HRP activity. **Procedure Summary:** 1. **Standard Dilution:** Prepare a serial dilution of the standard from 1800 ng/L down to 150 ng/L. 2. **Sample Loading:** Add 40 µL of sample diluent and 10 µL of sample into each well (final dilution 5×). 3. **Incubation:** Seal the plate and incubate at 37°C for 30 minutes. 4. **Washing:** Wash the plate 5 times with diluted washing buffer. 5. **Enzyme Conjugate Addition:** Add 50 µL of enzyme-labeled reagent to each well. 6. **Second Incubation:** Repeat the incubation step. 7. **Color Development:** Add 50 µL of TMB A and B, incubate at 37°C for 15 minutes. 8. **Stop Reaction:** Add 50 µL of stop solution to each well. 9. **Measurement:** Read the OD values at 450 nm within 15 minutes of stopping the reaction. **Important Notes:** - Allow the kit to equilibrate at room temperature before use. - Avoid cross-contamination by using a new sealing film for each test. - Always prepare a standard curve and run samples in duplicate. - Handle all reagents carefully, and dispose of waste as biohazardous material. - Follow the manual strictly; results must be based on instrument readings. **Performance Characteristics:** - Linear range: 0.2 IU/L – 6 IU/L - Correlation coefficient (R²) ≥ 0.95 - Intra-assay CV < 9%, Inter-assay CV < 11% **Technical Support:** We offer free technical support during working hours. Please contact us for assistance with sample testing or experimental design to ensure optimal results. **Disclaimer:** This kit is intended for research use only. Not for diagnostic or therapeutic purposes.

Silicone Pet Supplies

Silicone Pet Food Mat,Silicone Pet Bowl,Silicone Pet Bath Brush,Silicone Pet Bowl Mat

Ningbo Honor Imp & Exp Co.,Ltd , https://www.honormaket.com