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Description
Mouse TNFb ELISA KitProduct Specification Usage Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High precision pipette and gun tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37 constant temperature box 4. Distilled water or deionized water Sample processing and requirements: Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4 overnight, then centrifuge at 1000g for 20
Product Specification
| Usage |
Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High-precision pipette and gun tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37℃ constant temperature box 4. Distilled water or deionized water Sample processing and requirements: Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4℃ overnight, then centrifuge at 1000×g for 20 minutes, and take the supernatant, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. Plasma: Collect the specimen using EDTA or heparin as an anticoagulant. Centrifuge the specimen at 1000 × g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be assayed or stored at -20°C or -80°C, but avoid repeated freezing and thawing. Tissue homogenization: Rinse the tissue with pre-chilled PBS (0.01M, pH 7.4) to remove residual blood (lysed red blood cells in the homogenate will affect the measurement results). Weigh the tissue and mince it. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1 g of tissue sample to 9 mL of PBS. The specific volume can be adjusted according to experimental needs and recorded. It is recommended to add protease inhibitors to the PBS) in a glass homogenizer and grind thoroughly on ice. To further lyse tissue cells, the homogenate can be sonicated or repeatedly frozen and thawed. Finally, centrifuge the homogenate at 5000 × g for 5-10 minutes, and the supernatant can be assayed. Cell Lysis Buffer: Gently wash adherent cells with ice-cold PBS, then trypsinize and collect cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash collected cells three times with ice-cold PBS and resuspend in 150-200 μL of PBS per 1×10^6 cells (it is recommended to add protease inhibitors to the PBS; if the cell count is very low, reduce the PBS volume appropriately). Disrupt the cells by repeated freeze-thaw cycles or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and remove the supernatant for analysis. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, avoiding repeated freeze-thaw cycles. Other biological fluids: Centrifuge at 1000xg for 20 minutes, remove the supernatant, and test. Pre-test preparation: 1. Remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Prepare the standard gradient working solution: Add 1 mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 200 pg/mL). Then dilute to the following concentrations: 200 pg/mL, 100 pg/mL, 50 pg/mL, 25 pg/mL, 12.5 pg/mL, 6.25 pg/mL, 3.125 pg/mL, and 0 pg/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 200pg/mL standard working solution into the first EP tube and mix thoroughly to make a 100pg/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves as a blank well; there is no need to pipette liquid from the penultimate tube. See the figure below for details. 3. Preparation of biotinylated detection antibody working solution: Centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute 15 minutes before use. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration with universal diluent (e.g., 10uL concentrate + 990uL universal diluent). Prepare and use immediately. 4. Prepare the enzyme conjugate working solution: 15 minutes before use, centrifuge the 100× concentrated enzyme conjugate at 1000×g for 1 minute. Dilute the 100× concentrated HRP enzyme conjugate to a 1× working concentration with universal diluent (e.g., 10 μL of concentrate + 990 μL of universal diluent). Prepare immediately. 5. Prepare the 1× wash solution: Dispense 10 mL of 20× wash solution into 190 mL of distilled water (concentrated wash solution removed from the refrigerator may crystallize; this is normal. Allow to stand at room temperature until the crystals have completely dissolved before preparing). Procedure: 1. Remove the desired strips from the aluminum foil bag after equilibration at room temperature for 10 minutes. Seal the remaining strips in a ziplock bag and return to 4°C. 2. Sample addition: Add 100 μL of sample or standard of varying concentrations to the corresponding wells. Add 100 μL of universal diluent to the blank wells. Cover with a film and incubate at 37°C for 60 minutes. (Recommendation: Dilute the sample to be tested at least 1-fold with universal diluent before adding it to the ELISA plate. This will reduce the impact of matrix effects on the test results. The sample concentration should be multiplied by the corresponding dilution factor when calculating the final sample concentration. It is recommended to run replicates for all test samples and standards.) 3. Add Biotinylated Antibody: Remove the ELISA plate and discard the liquid without washing. Add 100 μL of Biotinylated Antibody Working Solution directly to each well. Cover with a film and incubate at 37°C for 60 minutes. 4. Wash: Discard the liquid and add 300 μL of 1x Wash Solution to each well. Let stand for 1 minute, shake off the wash solution, and pat dry on absorbent paper. Repeat this process three times (a plate washer can also be used). 5. Add Enzyme Conjugate Working Solution: Add 100 μL of Enzyme Conjugate Working Solution to each well. Cover with a film and incubate at 37°C for 30 minutes. 6. Washing: Discard the liquid and wash the plate five times as in step 4. 7. Adding substrate: Add 90 μL of substrate (TMB) to each well, cover with a sealing film, and incubate at 37°C in the dark for 15 minutes. 8. Adding stop solution: Remove the ELISA plate and add 50 μL of stop solution directly to each well. Immediately measure the OD value of each well at a wavelength of 450 nm. Calculating experimental results: 1. Calculate the average OD value of the standard and sample replicates and subtract the OD value of the blank well as a correction factor. Plot the standard curve of the four-parameter logistic function on double-logarithmic graph paper, with concentration as the horizontal axis and OD value as the vertical axis. 2. If the sample OD value is higher than the upper limit of the standard curve, dilute the sample appropriately and retest. Multiply the sample concentration by the corresponding dilution factor. |
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| Theory | This kit uses a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA). Sample, standard, biotin-labeled detection antibody, and HRP conjugate are sequentially added to microwells pre-coated with a Tumor Necrosis Factor Beta (TNFb) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB converts to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of Tumor Necrosis Factor Beta (TNFb) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Tumor Necrosis Factor Beta ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Tumor necrosis factor-β (TNF-β), also known as lymphotoxin-α (LT-α), is a cytokine-like protein encoded by the LTA gene. It is expressed in hematopoietic cells, exhibiting antiproliferative activity and leading to cytotoxicity in tumor cell lines. As a cytotoxic protein, it plays a variety of important roles in immune regulation, depending on the form in which it is secreted. Unlike other members of the TNF superfamily, it is found only as a soluble homotrimer and, when expressed on the cell surface, only as a heterotrimer with LT-β. It has a significant impact on the maintenance of the immune system, including the development of secondary lymphoid organs. LT-α deficiency leads to disrupted gastrointestinal development, prevented Peyer's patch development, and splenic disorders. It is involved in the regulation of cell survival, proliferation, differentiation, and apoptosis. Its effects vary depending on the organ type, cancer cell type, cellular environment, sex, and duration of action during the immune response. Produced by Th1 cells, it induces changes in adhesion molecules on the surface of vascular endothelial cells and promotes their binding to phagocytes. | |||||||||||||||||||||||||||||||||
| General Notes | 1. Strictly adhere to the specified incubation time and temperature to ensure accurate results. All reagents must be at room temperature (20-25°C) before use. Refrigerate reagents immediately after use. 2. Improper plate washing may result in inaccurate results. Ensure that all liquid in the wells is aspirated thoroughly before adding substrate. Do not allow the wells to dry out during incubation. 3. Remove any residual liquid and fingerprints from the bottom of the plate, as this will affect the OD value. 4. The substrate developer solution should be colorless or very light in color. Do not use substrate solution that has turned blue. 5. Avoid cross-contamination of reagents and specimens to prevent erroneous results. 6. Avoid direct exposure to strong light during storage and incubation. 7. Do not expose any reagents to bleaching solvents or the strong fumes emitted by bleaching solvents. Any bleaching agent will destroy the biological activity of the reagents in the kit. 8. Do not use expired products, and do not mix components with different product numbers and batches. 9. Recombinant proteins from sources other than the kit may not be compatible with the antibodies in this kit and will not be recognized. 10. If there is a possibility of disease transmission, all samples should be managed properly and samples and testing devices should be handled according to prescribed procedures. |
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| Storage Temp. | If the unopened kit is stored at 4°C, the shelf life is 6 months. | |||||||||||||||||||||||||||||||||
| Test Range | 3.12-200 pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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Product Reviews
★★★★★ 5
Perfect for Platform Beds and Pet Owners Perfecto para camas de plataforma y dueños de mascotas
Color: 1 Gray, Size: King
"This is a great option if you have a platform bed. It looks very beautiful as a bedspread and stays perfectly smooth. It's also perfect if you have pets because it prevents accidents. It’s very useful and looks great."
"Es una excelente opción si tienes una cama de plataforma. Se ve muy bonito como colcha y queda muy estiradito. Además, es perfecto si tienes mascotas porque evita accidentes. Me fue muy útil y se ve muy bien."
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Reviewed in the United States on April 22, 2026
★★★★★ 5
Utopia bedding is GREAT
Color: White, Size: Full
I had never heard of Utopia before I suddenly needed to buy bedding for my adult autistic son. He has sensory issues so shopping for anything that touches his body is a bit of a nightmare. I took a gamble on these sheets but it paid off big time! He says they are the most comfortable sheets he has ever slept on and that he doesn't overheat sleeping in them. What I liked about them is that the pockets were deep enough that it fit on his oversized plush mattress easily. I love the color and the ones I bought were very true to the picture. These are soft, comfy, they look nice and they are affordable. I liked this brand so much that I went ahead and got pillows and a comforter from them as well. Its is great for my son and I am very pleased to have found a fabric he tolerates.
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Reviewed in the United States on April 19, 2026
★★★★★ 5
Wonderful sheet set
Color: Grey, Size: California King
A very beautiful sheet set at an excellent price—the material is soft and comfortable! I loved the color!!!
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Reviewed in the United States on May 18, 2026
★★★★★ 5
Perfect Fit and Great Quality Sheet Set (10/10)
Color: Grey, Size: King, Color: Grey, Size: King
⭐⭐⭐⭐⭐⭐⭐⭐⭐⭐ (10/10)
This bedsheet has been working flawlessly and honestly exceeded my expectations.
The fit is perfect on the mattress—it wraps securely without slipping or coming loose during the night. It stays in place even after movement, which makes a big difference in comfort and convenience.
The fabric feels soft, comfortable, and well-made, giving a clean and fresh look to the bed. It’s also very easy to put on and remove, which makes washing and changing sheets much simpler.
So far, there are absolutely no issues at all. Everything works exactly as it should, and it holds up really well with daily use.
Overall, it’s a simple but high-quality product that does its job perfectly. I would definitely recommend it—10/10.
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Reviewed in the United States on May 10, 2026
★★★★★ 4
super comfortable, not waking up as often as before
Color: Grey, Size: Queen
decided to try these after seeing the tons of good reviews and since they were on sale. also one of my favorite colors in general which is a great neutral shade for bedding, not too bright and not too dull. I ordered a queen thinking my mattress might have been a queen as I previously bought some mainstays walmart sheets. with that brand the fitted sheet in full was way too small. turns out my mattress is in fact a full it is just that mainstays is not good quality or sized right, so this brand the queen is a bit big for my mattress but not too big of a deal. i am not looking for a perfect look, just covering the mattress is the most important.
anyway since getting these sheets in the mail and giving them a run through the wash and dry before putting them on my bed my sleep quality has improved. the sheets are not too hot, are soft and I actually do not wake up as often as before. the sheets are not too thin like other brands and are not thick like flannel either. they are a nice in between. I wonder if they will help keep me cool in the summer, probably since they are actually breathable. I would definitely reccommend giving these sheets a try, worth the money and a good deal on sale. better quality that other store bought sheets.
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Reviewed in the United States on March 16, 2026