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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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4.5 ★★★★★
Based on 5 reviews
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Product Reviews
★★★★★ 4
Amazing pillows!
Color: White, Size: Queen(Pack of 2)
These are fantastic pillows! A little background. I'm a side sleeper and toss and turn throughout the night. I've struggled with sleep issues for literally decades - insomnia, tossing and turning, waking up multiple times a night to use the bathroom, waking up way too early and not being able to get back to sleep. Every night it's a combo of those things or all of those things. I've also had some neck pain and stiffness for a couple of months. If you've ever dealt with that you know it will affect sleep which will then affect productivity and mood. It's a terrible spiraling and compounding thing to deal with, very stressful. I figured the contour pillow I had which once was actually helping was unfortunately the culprit. So I went on the hunt for another pillow. I ordered what I thought was the old Rejuvenite pillow (those were fabulous!) and got burned on a bait and switch. The pillow ended up being way too soft and gave me worse neck pain. Figuring I'd try to salvage what was a $115 pillow, I invested in a very thin pillow to put underneath. It was too high. Getting pretty frustrated, I ordered the cheaper Utopia pillows here on Amazon. Those actually were not terrible because they did take away my neck pain, but I was still tossing and turning at night and still feeling unrested in the morning. I decided I'd give it one last shot and luckily found these when they were discounted by a few bucks. They didn't have many reviews which was actually a great sign for me because it means the company lets their product speak for itself.
I got these in several days and was thrilled to see upon opening the package that they were sealed and the directions do not require you to open the package and let them expand for 24 hours. You literally just take the edges and fluff 2-3 times and that's it! They are ready to go and they definitely do feel perfect after doing that. I was skeptical when I put them on my bed because they are soft and I worried they'd be too soft. I felt my head sink into the pillow a bit and then was surprised to feel at a certain point it just feels like the pillow is cradling your head with the right amount of support. I wouldn't describe it as firm, just soft but stable support. Really comfortable.
But the real test was how it would perform during sleep. I'm so thrilled to report that I had an amazing night of sleep! No neck pain, no stiffness, zero tossing and turning, no insomnia, no waking up during the middle of the night, and no waking up early. I put my head down, fell asleep, and slept like a log for about 7.5 hours straight, which is literally unheard of in my world. I'm very lucky if I get 5 solid hours of sleep. I woke up and popped out of bed. My mood was great (still is later in the day) and I was not tired in the afternoon when I normally feel the fatigue kicking in. This is the first day for as long as I can remember that I do not feel that nagging tiredness behind my eyes and in my body that always seems to be a steady background actor in my life. I feel great and full of energy and I was able to tackle a bunch of tasks on my someday list. This is so promising I didn't want to wait to share. If you are on the fence and are dealing with similar issues to mine, I'd encourage you to just try these.
What's also amazing is these come in a set. I'm only using one so I basically have another one to use if the first one ever fails. This is the best investment for sleep I've ever made!
Update:
Well, after a trial period I'm removing a star because my dreamy sleep was short lived. I started to wake up more frequently during the night with these and gave up midway through the night one night. I'm still using these for back support while I sit up or hang out in bed because they are still super comfortable that way and oddly these work well for naps. Hopefully, you have better luck than me!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 21, 2026
★★★★★ 5
Actually feels like a hotel stay
Color: White, Size: Queen(Pack of 2), Color: White, Size: Queen(Pack of 2)
I've been looking for that specific hotel feel without spending a fortune, and these Serta pillows nailed it. They are categorized as medium-firm, which I find perfect, they have that initial plush softness when you lay down, but they don't flatten out completely by morning.
The 400-thread-count cotton cover feels high quality and breathable, so I haven't dealt with any overheating. I'm a side sleeper and my husband sleeps on his back, and we both find them super comfortable. If you want a pillow that balances support with that cloud like luxury these are a great pick.
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Reviewed in the United States on April 23, 2026
★★★★★ 3
Decent
Color: White, Size: Queen(Pack of 2)
They are only used as a backup pillow for my special pillows I use since I have pillows with holes in the middle for my earrings. I also use them for when I lean on my headboard to watch tv. I like them they are squishy. Kind of expensive I feel like but not terrible
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Reviewed in the United States on May 25, 2026
★★★★★ 5
I love these pillows
Color: White, Size: Queen(Pack of 2)
Very good pillows, comfortable and with that hotel style. They have medium firmness, so they work well for any sleeping position. They feel fresh, of good quality and do not crush easily.
Excellent purchase.
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Reviewed in the United States on April 20, 2026
★★★★★ 5
Worth it, I have 2
Style: All Season, Size: King
Soft, good material, I like the thickness. Does not feel cheap.You do have to put it in a dryer on really low to fluff it up. Check it after 20 minutes. Pull it out fluff it up and put it back in the dryer. Do not use hot heat. I also put it in a linen duvet.
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Reviewed in the United States on May 1, 2026