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Description
Rat IL-35 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 1mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 1000pg/mL). Then dilute to the following concentrations: 1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.25pg/mL, 15.625pg/mL, and 0pg/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 1000pg/mL standard working solution into the first EP tube and mix thoroughly to make a 500pg/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 utilizes 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 an IL-35 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 IL-35 content in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat IL-35 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Interleukin-35 (IL-35) is an anti-inflammatory cytokine of the IL-12 family. As a member of the IL-12 family, IL-35 is produced by a wide range of regulatory lymphocytes and plays a role in immunosuppression. IL-35 can inhibit the development of Th1 and Th17 cells by limiting early T cell proliferation. IL-35 is a dimeric protein composed of the IL-12α and IL-27β chains, encoded by two separate genes, IL12A and EBI3 (Epstein-Barr virus-induced gene 3), respectively. Its receptors consist of the IL-12Rβ2 (part of the IL-12R) and gp130 (part of the IL-27R) chains. In contrast to these two related interleukins, IL-35 can signal through only one of these chains. This has been demonstrated in vivo, as the absence of either receptor chain does not affect the effects of IL-35. On regulatory B cells, IL-35 signals through the IL-12Rβ2 and IL-27Rα subunits. | |||||||||||||||||||||||||||||||||
| 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 | 15.6-1000 pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.2 ★★★★★
Based on 1400 reviews
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Product Reviews
★★★★★ 5
Great mouse
Number of Items: 1
Definitely reduces fatigue. Easy to use. Great response with less space needed for mouse movement. Stays charged for months you actually forget to charge it, ask me how I know :). I play video games with it and never have any issues with the movement and response time of mouse. And since my car wreck where I broke my thumb this is the only mouse that gave me the same as before the wreck abilities to move and operate the mouse. Highly recommend. One very broken thumbs up :)
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Reviewed in the United States on May 31, 2026
★★★★★ 4
Experience after one year of usage
Number of Items: 1
I had mainly used a MX Master 2S before this and because of my heavy usage I did start to feel uncomfortable after a few hours of use every day and that's when I decided to purchase MX Vertical. Now after almost 9 months of usage this has become the mouse I use on a daily basis, completely replacing MX Master 2S.
Comfort: It took me about a day or two to really get used to using it because of the positioning and ever since then my wrist has never felt any discomfort, literally. I don't have a mouse pad but because of the angle that my wrist is contact with the desk when I use it, this has never been a problem. What I also love about the ergonomic design is that my hand can actually just rest on it when not doing specific tasks, and it just fits my palm perfectly.
Functionality: I don't use it for gaming so I can't comment on that. However, comparing it with my Master 2S, there are some very obvious disadvantages. If you use Excel sheets a lot, or anything that requires horizontal viewing, the scrolling option is terrible. You can set up the function key on Logi Options+ and I use the side buttons for scroll right+left mainly because how easy it was to use with the Master 2S's side scrolling buttons. It is not comparable at all. With Master 2s, horizontal scrolling is smooth and precise. With this, it's basically useless. There are sites that just aren't responsive to the side buttons and when they do, you can never get to the precise point you want. Huge disappointment with that because I don't really use the side buttons for anything else. There is also no scrolling control with the middle button unlike the Master 2S, so you can't really scroll vertically as fast as you can when using Master 2S either. However I do use the middle button sometimes to just do the job for me so I'm ok with its basic functionality.
The top button is mainly used as a look-up button for me, so far its working pretty well and honestly it probably makes not much difference if they just completely take this button out for me. Maybe some people can find use in it as a switch for scrolling or mouse speed, or something else. Overall, the customizability of these buttons is very good, and depending on personal use, I can see it being very useful.
Battery life: Battery life is fantastic, never had any problem with it yet. I haven't really kept track but with my heavy daily usage (probably 10+ hours with no intense gaming), it can usually go for more than a month. You can also use it while it's being charged.
Durability: I have unfortunately knocked it over the desk a couple times on hard vinyl floor and there seems to be no problem with it at all. Before purchasing this I saw some comments about how it stopped functioning after being dropped to the floor and I was really worried about it. However that is not my case at all. It is very durable in my opinion and does its job perfectly even after dropping it a few times.
Overall, extremely comfortable to use. It helped with my wrist pain tremendously and now I use it for hours without feeling any discomfort. The functionality is severely disadvantageous compared to Master 2s and 3s, and possibly other gaming mouses as well, but if you are here because of wrist pain, this is the right product.
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Reviewed in the United States on May 24, 2024
★★★★★ 5
Finally, a comfortable vertical mouse for guys with large hands!
Number of Items: 1
I've been using the Logitech MX Vertical Wireless Mouse for about 8 years now, and I love it. Any time I have to use a traditional mouse it's an uncomfortable disappointment.
If you haven't used a vertical mouse yet, there are a couple things you need to know.
First - this is just a normal mouse, but it's turned about 60-degrees to the right. This let's your hand rest on the mouse in the natural position your arm wants it to be in without any effort to rotate.
Second - you will knock it over, repeatedly, for a few weeks. Because you're used to a mouse that's a just an inch and a half or so tall, your natural movement from keyboard to mouse is going to be interrupted by the height of the MX Vertical. In time, you will get used to this, but for a week or two, you will knock it over when moving to the mouse, and when moving back to the keyboard. Don't worry, your muscle memory will change and this will stop.
Third - once you get used to using the MX Vertical, you won't want to use any other mouse. It's a dramatic change in ergonomics, and you will feel the strain your wrist and forearm are under any time you have to use a traditional mouse.
There are other verticals that are also great, but as a guy with large hands, this is the best I've found. I have 4 of these now - one at my office, one at my dining room table, one at my desk, and one that travels in my backpack. I don't need all of these, but its handy having them where I need them. Another reason for this is the mouse can pair with 3 devices. I have different devices in each of these locations, so it's handy not having to re-pair when I move between locations.
The MX Vertical fits my hand perfect, and all buttons are where my fingers and thumb are naturally positioned. I've dropped them, knocked them down, bumped and tossed them and they keep on working properly, so from a durability standpoint, this will last. It's easy to pair with a push of a button - working with my Windows computers, MacBook, android phone and iPad. This thing just works great!
The only downside I'd mention is the coating. After a couple of years, the coating tends to migrate where you apply pressure. This is most apparent where the thumb and pinky finger rest. This doesn't cause any functional issues, and is entirely cosmetic, but just be aware that after a couple years it won't look pristine.
All-in-all, this is the vertical mouse to buy if you're a guy with larger hands. Get one, get used to it, and you won't regret it.
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Reviewed in the United States on February 6, 2026
★★★★★ 5
Highly recommend! Best in the market bar none
Number of Items: 1
I would like to say this is the perfect mouse and probably the best mouse you could buy on the market and it doesn't even come close. You can modify the mouse however you like with a click on the app. It's truly been a life changing experience with this mouse. With that being said, the perfect mouse does have a few flaws but nothing that would make me not recommend it to anyone only just be aware of these things. 1. It's a vertical mouse, sometimes it gets in the way and your wrist will knock it over, it will happen I promise you. 2. Logitech has a Flow feature meaning you can connect the mouse to three different computers at once if need be, qyeah I'll say it again, three computers connection! Wow! Well I only use it for two computers, the Flow works flawlessly but my gripe is how it sometimes doesn't sync with my Logitech keyboard Ergo K860, unfortunately, the keyboard doesn't come with the same Flow feature meaning I have to depend on the mouse to do it and it doesn't work sometimes and I have to switch between the mouse transition button to the key board transition button which makes for a long headache sometimes. 3. Believe it or not connection is sometimes an issue, it losses Bluetooth connection often and it doesn't matter which computer I use I have the same issues at work and home so I use the dongle for straight connection. Oh wait there are issues with the dongle connection too? Say it ain't so? It is! The issue comes when you have two dongles like me, the keyboard and the mouse, and try to connect both, like I said I have two computers and Logitech will not allow for their ”universal dongle” to connect to two different computers at the same time terrible design flaw. 4. Again, this mouse is incredible but it can sometimes be a little too much incredible, I know, I know but hear me out. The mouse is flawless fast, one quick wrist jerk and that mouse pointer on the screen is jumping all over the place, it's been hard to find a happy medium with the pointer, it's either you adjust to slow it down, making it too slow, or adjust to make it fast, which is too fast, again no happy medium even though there are plenty of settings to toggle with. I believe most vertical mouse tend to be that way anyway only with this mouse you can really tell the difference and that could be a good thing or bad thing depending on your perspective, it's sometimes a bad thing for me on Excel sheets. So the final verdict is this is still the best mouse out there and worth every penny and I would still give this 10 stars if possible, it's been a wonderful experience with this mouse. Again, the negatives are not bad or to make someone not buy it and considering the other features this mouse can do I recommend anyone who works all day on a computer to buy this product I highly recommend it.
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Reviewed in the United States on July 28, 2021
★★★★★ 5
Excellent Ergonomic Mouse for Long Workdays
Number of Items: 1
If you spend many hours at a computer and are looking for a comfortable, high-quality mouse, I highly recommend the Logitech MX Vertical. It has made a noticeable difference in my daily comfort and productivity.
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Reviewed in the United States on June 5, 2026
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