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Description
Rat TNF-α 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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| Sensitivity | 1.75 pg/mL | |||||||||||||||||||||||||||||||||
| 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 Alpha (TNF-a) 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 Alpha (TNF-a) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Tumor Necrosis Factor Alpha ELISA Kit，Rat TNF-a ELISA Kit，TNF-a | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Tumor necrosis factor alpha (TNF-α) is produced by neutrophils, activated lymphocytes, macrophages, natural killer (NK) cells, lymphocyte kinase (LAK) cells, astrocytes, endothelial cells, smooth muscle cells, and some transformed cells. The naturally occurring form of TNF-α is glycosylated, but nonglycosylated recombinant TNF-α exhibits comparable biological activity. The biologically active native form of TNF-α is reported to be a trimer. Human and murine TNF-α exhibit approximately 79% homology at the amino acid level, and cross-reactivity is observed between the two species. | |||||||||||||||||||||||||||||||||
| 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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4.1 ★★★★★
Based on 379 reviews
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Product Reviews
★★★★★ 5
Finally a Cooling Blanket That Actually Works for Hot Sleepers
Size: Queen (90" * 90"）, Color: Luxury Grey, Size: Queen (90" * 90"）, Color: Luxury Grey
I first got to know this brand when they released their cooling pillow. At the time, I was very skeptical because I’ve tried many “cooling” pillows and pillowcases from other brands, and almost all of them failed to stay cool throughout the night. Surprisingly, this brand was the first one that actually worked for me.
Going through menopause has been extremely difficult, especially dealing with the constant heat and night sweats. I kept thinking that if they could somehow use the same cooling technology in a blanket, it would be amazing — and they actually did it.
This blanket has honestly been a lifesaver for me. One side has the cooling fabric while the other side feels more like a regular blanket, so you can switch depending on the temperature or season. I’ve been using a silk comforter before this, and while it was comfortable, it really didn’t help much with overheating at night. This cooling blanket made a noticeable difference almost immediately.
What impressed me most is that the cooling effect doesn’t just disappear after a few minutes. It stays comfortably cool and lightweight through the night without feeling damp or overly thin. Since using it, I’ve actually been tracking improvements in my sleep using my WHOOP device, and over the past 20 days I’ve noticed my deep sleep readings improving consistently.
The blanket also feels well made, soft, and lightweight without feeling cheap. For the quality and performance, I think the price is very fair, especially compared to other “cooling” bedding products that overpromise and underdeliver.
If you struggle with hot sleeping, night sweats, or menopause-related heat issues, I would absolutely recommend giving this a try. It genuinely improved my sleep quality.
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Reviewed in the United States on May 24, 2026
★★★★★ 5
Great blanket for keeping cool, but doesn't magically defy physics.
Size: King（103“*90”）, Color: Luxury Grey
This blanket is pretty awesome, and functions exactly as stated; however you might get the wrong impression from the image of the blanket in a the snowy mountains glowing blue as if its actively cooling. The blanket has two sides, the "cold" and "normal" side -- the cold side is the one that has the Q-max 0.45 effect, it actually feels cool to the touch. That being said, covering yourself with this blanket feels pretty similar to using a down blanket with a silk (or some cooler) fabric. It feels quite cool initially but eventually your body puts out enough heat and it "runs out" of cooling.
Although this blanket doesn't magically actively generate cooling, its quite effective because it operates fundamentally differently from most blankets. Most blankets are engineered to keep you warm by trapping heat in; this blanket is explicitly designed to draw heat away from your body. As such, it is extremely effective if you use a fan or run the AC, because your body heat is drawn away from the top surface and the inner surface can "grab" and remove more heat. As you probably know, air is a great insulator so this blanket has to be making contact with your body to achieve the cooling effect. If you are familiar with the way "Under Armor" works, I believe this blanket uses a similar principle to keep you cool.
This blanket is quite light weight and comfortable, with a very silky (polyester) feeling outer surface. I have not owned this for a year yet, but I'm quite confident that this blanket would be suitable for winter (in Southern California); except if you live in a snowy region.
5/5 for a somewhat high tech blanket that keeps you cool; its the best in cooling you can get without resorting to "active" cooled blankets such as ones with water circulation.
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Reviewed in the United States on May 5, 2026
★★★★★ 5
Fascinating product.
Size: Twin（90“ * 67”）, Color: Luxury Grey
It is a nightly struggle. My wife is a hot sleeper. I am not. It’s resulted in more than one argument. One of us is usually left shivering or sweating. So when Amazon offered to send me this cooling blanket, I thought I would give it a try. I’m really glad that I did.
This item arrived well packaged and in good condition. It is lightweight and made of a polyester type material. The stitching is strong. The first test was the washing machine. We washed it with cold water on the gentle cycle. We then line dried it. It held up fine.
We have a king size bed, but I decided to get a twin sized blanket. I put a warm blanket on my side of the bed and the cooling blanket on her side of the bed. It worked brilliantly. I don’t know what the science is buying these blankets, but it really does lower your body temperature. The moment that you open the package and feel the blanket, you’ll realize that it has some sort of cooling gel inside of it or something that draws heat away from you.
Suffice it to say that our first night with this product was one of the best nights sleep that both of us had. If you have a problem like this, I would definitely recommend this blanket as a viable solution.
In terms of value, I understand that this is not cheap; however, ask yourself what the value is of a good nights sleep. Overall, I’m very pleased with the quality, design, and functionality of the cooling blanket. I would definitely recommend it.
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Reviewed in the United States on May 5, 2026
★★★★★ 5
Cooler than my thinner blankets
Size: Queen (90" * 90"）, Color: Luxury Grey
This is a really nice cooling blanket—silky, soft, and comfortable. I actually tried it on a cold day (classic flip flip Rocky Mountain weather where it swings from 90° to 30° overnight), and it was absolutely freezing, lol. On warmer days, I can clearly tell that it's cooler than even my thinner blankets. My husband tends to sleep hot, and this works great for him. I run cold; it can feel a bit too cool initially, but that fades after a few minutes and stays comfortable.
I also have two cats and was worried they’d snag it right away. We keep their nails trimmed, but they’re still cats. So far (about a week in), no snags or pulls—but I’ll update if that changes since that’s a big factor for me.
At almost $100, it’s definitely not cheap for a blanket. I think it makes the most sense if you’re a hot sleeper or deal with night sweats—then the cooling effect feels more worth it. If you tend to run cold, it might feel like overkill unless you specifically want something for summer. Overall good value for a nice quality cooling blanket.
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Reviewed in the United States on May 4, 2026
★★★★★ 5
Surprisingly strong cooling effect and really comfortable
Size: Queen (90" * 90"）, Color: Luxury Grey, Size: Queen (90" * 90"）, Color: Luxury Grey
This comforter honestly impressed me more than I expected. The cooling effect is very noticeable right away. Like actually noticeable, not just one of those things brands say. When you first touch it, it feels cool to the skin and that feeling sticks around instead of fading instantly.
I’ve been using it at night and it definitely helps with overheating. I tend to get cery hot when I sleep, also I’m pregnant right now so that makes it even worse, and this has made a difference. It feels lightweight but still cozy enough that it doesn’t feel like you’re just using a thin sheet.
The fabric is really soft and smooth, also it lays nicely on the bed without feeling stiff or bulky. It gives that clean, almost silky feel without being slippery.
One thing to keep in mind is it’s not a super thick comforter, but i will say there is some heft to it. Like if i wrap myself around in it, it has some weight to it.
Overall, I’m really happy with it. It actually does what it claims with the cooling, and it’s comfortable enough for everyday use. Great option if you’re a hot sleeper or just want something lighter and refreshing at night.
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Reviewed in the United States on April 22, 2026