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Description
Mouse IL27 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: 1. 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: 1. 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. 2. 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. 3. 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. 4. 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. 5. 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. 6. 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 500pg/mL). Then dilute to the following concentrations: 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.25pg/mL, 15.625pg/mL, 7.8125pg/mL, and 0pg/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each tube. Pipette 500uL of the 500pg/mL standard working solution into the first EP tube and mix thoroughly to make a 250pg/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 Interleukin 27 (IL27) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by peroxidase (HRP) catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of Interleukin 27 (IL27) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Interleukin 27 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Interleukin-27 (IL-27) is a heterodimer composed of EBI3 and p28 subunits linked by disulfide bonds. IL-27 promotes Th1 and Tr1 differentiation and inhibits Th2, Th17, and Treg responses, playing a crucial role in immune regulation. IL-27, a member of the IL-12 family, is primarily produced by antigen-presenting cells, including dendritic cells and macrophages, in response to microbes or other immune stimuli. Its receptors are widely distributed on various immune cells. IL-27 signals are transmitted intracellularly through the heterodimeric receptor IL-27R, which is located on cells. IL-27R is composed of WSX-1 and gp130 subunits. WSX-1 and gp130 are expressed on a variety of cells, including T cells, NK cells, B cells, and myeloid cells. | |||||||||||||||||||||||||||||||||
| 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 | 7.81-500 pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.3 ★★★★★
Based on 11 reviews
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Product Reviews
★★★★★ 5
Unexpensive and great for summer
Size: King, Color: White
Light, synthetic, but oeko tex label.
White, breathable, easy to wash and dry, comfy, seems a great quality. It will last for a while.
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Reviewed in the United States on January 19, 2026
★★★★★ 5
A 5 star WIN - such a soft, so fluffy, perfect cloud duvet comforter that feels expensive but isn't. Like sleeping in a cloud.
My fiance' and I both had king size duvets - mine from Ikea, down alternative, a few years old, and I'm not sure where his was from, but his was true goose down/feathers. After going away for the weekend to stay at a lovely vacation home, we got a bit spoiled with the comfortable bed and great linens there, so when we came home we decided it was time for a bed makeover. Since I love to research, I spent hours scouring Amazon for something that looked like it was actually a great comforter at a great price.
This duvet is everything we wanted and more. Oh my GOSH...it is SO fluffy. SO soft. It's my marshmallow cloud comforter. The inside fill has a silky soft feeling to it when you press on it - it sort of slides in your hands when you touch it, like the fill has a light, silky feel to it Even when I sit on the end of the bed to do something, I can feel how comfortable it is to just even sit on. It makes our old duvets seem pathetic, like something you'd go camp with. This duvet is as nice as the bedding we had where we stayed in our vacation home.
It feels like it has medium weight, but it's warm - quite warm for the weight. We keep it cool in our home at night so for us it's perfect. The material is so soft. We covered it immediately with a beautiful cover because we want to keep it in perfect condition.
I also notice that even though both of our old duvets were king size, this one (also a king) just seems bigger. Our bed is a queen and the king size fits perfectly over it, it drapes over the sides just far enough. Also, my fiance' likes to roll up in his covers when he sleeps. He has plenty of room to do that without taking any of the duvet from my side. :)
Verdict - it's amazing, I feel like a princess sleeping under this duvet. It's amazing for this price, amazing. All 3 of us give it 5 stars - me, fiance' and our cat who now won't get off of our bed.
Someone mentioned that it doesn't have loops on the corners. That's true, it doesn't, but you can always sew those on if you want to. I found a kit for that on amazon too. But trust me, this comforter is worth sewing loops if you really want them. However, like others, we haven't found that the comforter moves or shifts in the duvet cover at all. It stays very well, better than any of my old comforters did, and I've had a few. Perhaps it's the loft that keeps it in place so well.
I'll check back in after some time to let you know if if stays nice.
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Reviewed in the United States on March 28, 2016
★★★★★ 4
Soft, Cozy, and Perfect for All Seasons – Great Value for the Price!
Size: King, Color: Grey
I purchased the Utopia Bedding All Season Down Alternative Quilted King Comforter, and it has been a great addition to our bed. The comforter is soft and has a nice, lightweight feel that’s comfortable year-round. It provides just the right amount of warmth without feeling too heavy, so we’ve used it both during cooler nights and warmer weather without feeling overheated. The grey color is rich and stylish, adding a modern touch to our room decor.
The quilted design keeps the filling evenly distributed, so there are no cold spots, and it holds its shape well even after washing. The corner tabs are a nice touch for securing it inside a duvet cover, though we’ve also used it on its own. It’s easy to wash in the machine, and so far, it’s held up well with no signs of pilling or fading.
One minor drawback is that it’s a bit thinner than I expected for a “down alternative” comforter, so if you prefer a super fluffy look, you might want to layer it with an additional blanket. That said, it’s perfect for a clean, streamlined appearance on the bed, and the comfort level is still very cozy.
Overall, this Utopia Bedding comforter is a fantastic value for anyone seeking an affordable, all-season option that’s easy to care for. It’s comfortable, versatile, and stylish – a solid choice if you’re looking for a dependable comforter for everyday use!
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Reviewed in the United States on November 2, 2024
★★★★★ 5
So great to use as a Duvet insert or as a Comforter on it’s own.
Size: King, Color: White, Size: King, Color: White
I recently purchased this Duvet Insert/All Season Comforter and I am thoroughly impressed with its performance. The bright white color seamlessly complements my furniture and walls, adding a clean and elegant look to my bedroom. What I appreciate most is its versatility; it keeps me warm during the winter months while also providing a cool comfort when the temperature rises.
I love that it's lightweight yet still offers that cozy, snug feeling, which is why I haven't felt the need to use a duvet cover. The initial setup was a breeze—I simply took it out of the shrink wrap, tossed it in the washer and dryer, and it was ready to use immediately. There was no unpleasant odor upon opening, which is a big plus.
The functionality of this comforter is commendable; it serves well both as a standalone comforter and as an insert for a duvet cover, making it a practical addition to my bedding collection. After washing and drying, it retained its fluffy shape and vibrant color, further enhancing my satisfaction with the purchase. Overall, it's a great product that combines style and comfort effectively.
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Reviewed in the United States on February 1, 2025
★★★★★ 5
Great duvet insert at a budget price
Size: Queen, Color: White
I'm new to using a duvet so I had no idea where to even start when it came to a duvet insert. I decided on this one and what a great decision that was. This duvet insert is perfect and fit my queen size cover perfectly. It did arrive vacuum sealed but once you cut it open it quickly regained its shape. I did throw it in the dryer for 10mins with no heat and that seemed to help add an extra layer of fluffiness but I do not think that is necessary for everyone. This duvet insert is perfect for all year around because it is not heavy, so it will keep you warm without getting too hot in the summer. Overall, I am very pleased with my purchase and I would highly recommend. Also, it is washable which is such a plus!
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Reviewed in the United States on August 28, 2024