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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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4.5 ★★★★★
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Product Reviews
★★★★★ 5
Fantastic for gaming and productivity, but missing USB-C rechargeability
Color: Classic Black, Color: Classic Black
I recently purchased the Razer Orochi V2, and overall, it is an amazing mouse. The build quality is absolutely fantastic, and it feels premium. It fits perfectly in my palm, and the materials and clicks have a very pleasant tactile feel. I use it daily for both gaming and work, and it performs flawlessly in both scenarios. Compared to my Razer Pro Click, the Orochi V2 actually fits my hand much better. In fact, I now only use the Pro Click as a backup for when the Orochi's battery suddenly dies!
There is really only one downside for me: it runs on disposable batteries rather than having a built-in rechargeable one. On the plus side, it only needs a single battery to operate, and I love the clever design that allows you to use either a standard AA or a smaller AAA battery depending on your weight preference.
However, I would really love to see a future version of this exact mouse with a built-in battery and a standard USB Type-C charging port. If you don't mind swapping batteries occasionally, this is a near-perfect mouse. Highly recommended!
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Reviewed in the United States on April 6, 2026
★★★★★ 5
Amazing mouse even for big hands.
Color: White
So I have big hands and what any big hand gamer understands is that you either have to get a big ol heavy G502 size mouse or use a certain grip to hold all these mice for smaller handed people. I previously owned a Model O, G pro, and Model O wireless.
Having big hands my biggest peeve with all the other mice is that they’re ambidextrous mice so they are even harder to hold with fingertip/claw grips because the smooth, near flat, symmetrical sides. I only really make contact with the sides by pinching my fingers with them. I never make contact with the butt or base of the mouse with my grip. There’s where the Orochi changes things
The Orochi somehow is a smaller mouse than all of the above but fills out my hand and grip SO much better. The sides actually have slight grooves so I don’t slip off the sides of my mouse and the shorter but bigger hump in the middle makes me for once actually feel the mouse in my palm if I want. Overall the feel is amazing.
Battery life varies of course but I can barely tell there’s a battery in the thing even when flicking and the weight and balance of it feel so good.
Mobile gaming is the main reason I got this mouse. I like that the dongle stores inside the mouse, uses batteries so I don’t have to worry about keeping it charged or bringing around extra cables to charge it, and the Bluetooth is good for casual gaming but doesn’t do great with big flicks so the 2.4ghz connection is required for esports games.
Overall a great mouse and really awesome to game with and a little plus to it is that the shell comes off the back so satisfying, it’s hard to not fidget with it by taking it on and off haha. Well worth the money and I’d expect they’ll make a rechargeable one with all the success this mouse has accrued so far so if the swappable batteries don’t tickle your pickle, I’d just buy a sub $20 rechargeable battery station or just wait for a new rechargeable Orochi to come out.
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Reviewed in the United States on May 20, 2021
★★★★★ 5
Unleash Your Gaming Potential
Color: White
The Razer Orochi V2 Mobile Wireless Gaming Mouse is nothing short of a gaming marvel. As a dedicated gamer, I've had the pleasure of using various gaming mice over the years, but the Orochi V2 stands out as a true game-changer. Here's why I can't help but sing its praises:
Wireless Freedom: The wireless connectivity of the Orochi V2 is a game-changer. With Razer's HyperSpeed Wireless technology, I experienced zero lag or interruptions during intense gaming sessions. The freedom to move without being tethered by a cable is liberating.
Compact and Portable: This mouse is designed with mobility in mind. It's incredibly compact and lightweight, making it perfect for gaming on the go. Whether you're a frequent traveler or just prefer a more minimalist setup, the Orochi V2 is the ideal companion.
Exceptional Battery Life: The Orochi V2 boasts impressive battery life. I was pleasantly surprised by how long it lasts on a single charge. It easily survived through extended gaming marathons and even lasted me multiple days of regular use without needing a recharge.
Customizable Performance: Razer's customization options are legendary, and the Orochi V2 doesn't disappoint. With the Razer Synapse software, you can fine-tune the mouse's sensitivity, button assignments, and lighting effects to suit your gaming preferences.
Versatile Connectivity: The Orochi V2 doesn't limit you to just wireless gaming. It also offers Bluetooth connectivity, allowing you to seamlessly switch between devices. This versatility is a godsend for those who use multiple devices for work and play.
Impressive Sensor: The 5G Advanced Optical Sensor delivers precise tracking and accuracy. Whether you're engaged in fast-paced FPS battles or navigating intricate RPG menus, the Orochi V2 responds flawlessly to your movements.
Comfortable Design: Despite its compact size, the Orochi V2 is surprisingly comfortable to use. It's designed to fit various grip styles, and the textured side grips ensure a secure hold during intense gaming moments.
In conclusion, the Razer Orochi V2 Mobile Wireless Gaming Mouse is a true gem for gamers who value performance, mobility, and customization. Its wireless capabilities, exceptional battery life, and versatility make it a standout choice for both casual and competitive gamers. Razer's commitment to quality and innovation is evident in every aspect of this mouse. If you're in the market for a compact and powerful gaming mouse that can keep up with your gaming lifestyle, the Orochi V2 should be at the top of your list. It has certainly elevated my gaming experience, and I couldn't be happier with this exceptional piece of gaming hardware.
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Reviewed in the United States on September 14, 2023
★★★★★ 4
Great quick mouse <3
Color: White
Very nice mouse though the scroll wheel is a little hard but it doesn’t skip around or behave oddly. instant set up and very comfy! I’m a girl with small hands so it fit perfectly! not too big or uncomfy, I can naturally grip it and the color is so perfect for my pastel blue and white setup! The click is a bit loud but not annoyingly loud, feels very sturdy despite it being so light, I’d say it’s a good price for a gaming mouse! (no connection issues either with Mac) I don’t have a mouse pad and it actually glides pretty good on my wooden desk.
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Reviewed in the United States on May 1, 2026
★★★★★ 5
Oh Boy
Color: Classic Black
Great design. Matte feel on the casing. Subtle logo on the black model (yeet). Nice and solid feel. The left/right clicks also feel premium and tactile
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Reviewed in the United States on June 2, 2026