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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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4.6 ★★★★★
Based on 7 reviews
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Product Reviews
★★★★★ 5
User friendly, Pet Hair Approved, Quite Powerful Robot Vacuum
Color: White
We prviously had an Eureka E10 vacuum and mop robot for 4 years. We had no idea what we were missing until we purchased this Narwal Freo X10 Pro.
It maps very fast.
It is extremely quite.
The vacuum then mop with a refreshed mop head solution is much better than dragging around a robot mop pad with no way to clean it without removing and washing it.
We have 3 Australian Sheppards that shed alot. This was the number one concern, whereas the prvious unit needed daily attention. Not a single hair tangle in the floating rotating brush of this unique Narwal Freo X10 Pro Robot vacuum.
4 months of no robot management is wonderful.
The application was easy to use with a ton of customizable options.
The AI option to recognize various floor types and throw rugs ensures a complete clean and quick simple setup.
Lastly the price vs product was very reasonable.
Great product! After spending many months researching, we are glad we made this investment.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on December 5, 2025
★★★★★ 1
After five months, it's basically a brick
Color: White
We've had this robot for five months, and it isn't durable. It worked fine for a couple months. But now it works fine for maybe 20 minutes and then just starts spinning around or going back in forth in the same area wasting its battery. The endless troubleshooting does not help. Cleaning all the sensors, resetting the robot, remapping the house ... Nothing helps. It is only being used in about 1100 sq ft of total space, so it's just not that complicated. It's been trying (and failing) to vacuum our smallish living room for nearly five hours, after several resets/etc. Back when it did function, the vacuum is rather loud if you want it to actually pick up anything big enough to see. The vacuum and mop are both terrible at inner corners, and the intersection between rugs and hardwood flooring is a total miss. There's like a 12" strip along all rugs where mopping never occurs. Definitely not a machine to be used without supplemental sweeping and mopping. The machine insists on starting rugs at the corner, so it pulls up even pretty heavy rugs sometimes. It's sucks up items like socks instead of avoiding them. Also, the machine has never successfully left alone no-vacuum rugs or no-go areas, so some rooms (like with high pile rugs or Christmas tree stands) just don't work without physical barricades. Really, really makes me miss the old reliable roombas...
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Reviewed in the United States on April 21, 2026
★★★★★ 5
This vacuum is a game changer
Color: White
Not all robotic vacuums are the same, and after testing so many of them in my life this one has outperformed the others hands down! From the suction power, to the battery life, and then the app I can't find a single thing I don't love.
The robot is and to pickup small crumbs, dirt, the typical messes, but most importantly it picks up kitty litter like it's nothing. This is an instant game changer. I no longer need to clean up after a robot. It's amazing!
The battery life is great! I have a 2500 sq ft house and it can handle all of it before charging. The "home base" makes this effortless where I just schedule my cleans and it takes care of the rest.
The app is really nicely built. It's simple and easy to navigate but it also gives me all of the advanced options such as invisible walls and no-go zones.
Overall, this is robotic vacuum is my new go-to and I look forward to using it for many years to come!
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Reviewed in the United States on April 14, 2026
★★★★★ 4
$500, not $600. It isn’t worth $600
Color: White
You’re gonna need to put on some glasses and grab a drink for this one.
Cons :
- Has issues with “muddy mats” brand rugs and mats, along with light mats
- It ate my socks
- Will molest your standing fans and shoes and pet toys
- Castor wheel is a hair magnet along with the “tangle less” floating brush
- Mop heads get dirty after just one run of the entire house
- It hates charging cables
- And dryer sheets
PROS:
- Smarter than Shark brand vacuums
- Excellent management of houses and avoiding most obstacles
- It mops and vacuums well
- Cleaning is very easy
- Cat litter gets sucked up on the strong setting (not the super strong, though that works too)
- Battery lasts for vacuuming most of the house
- No navigation issues
- Doesn’t mess up your furniture
- IT WORKS IN THE DARK
the floating brush though where the metal shaft sits inside the brush itself and the miniscule space between the spinning part and the brush itself gathers hair like a magnet and will give constant roller brush errors if not regularly cleaned. The castor wheel I swear has more hair than anything I’ve seen. Even the shark didn’t collect it this bad.
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Reviewed in the United States on April 17, 2026
★★★★★ 5
Value for Money- Exceptional performance and features!!!!
This vacuum/mop system is wonderful! I highly recommend it. This is bi-far the best vacuum and moping system which effectively cleans my floors and carpet. I've always been on the lookout for anything that can lighten my cleaning load.
it’s been a real game-changer for my household routine. The self-washing and self-charging features are a huge win. I no longer have to worry about cleaning the mop or making sure it’s charged; it takes care of itself.
It moves around furniture and other obstacles smoothly, rarely bumping into anything. It easily navigates tight spaces and corners, making sure every area gets cleaned.
Value for Money: Considering all its features and how well it cleans, I think it’s a fantastic investment. It’s made keeping my home clean so much easier and less time-consuming.
The robot vacuum was able to map my house successfully, the battery life is a bit longer than my previous robot vacuums, the suction is also great. I saw it go over crumbs in the kitchen and it picked up without any problems, and also has a mop functionality. It has a functioning app, very easy to use. The vacuum is also very quiet. By far, this is my favorite robot vacuum and I’ve tried many as I do not like to go around the house mopping and vacuuming. 10/10 would recommend this model - worth the investment.
Overall I could not be happier with this purchase, my home is the cleanest it has been since I’ve first moved in.
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Reviewed in the United States on May 12, 2025