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Description
Mouse IL-33 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 1 mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 400 pg/mL). Then dilute to the following concentrations: 400 pg/mL, 200 pg/mL, 100 pg/mL, 50 pg/mL, 25 pg/mL, 12.5 pg/mL, 6.25 pg/mL, and 0 pg/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each tube. Pipette 500uL of the 400pg/mL standard working solution into the first EP tube and mix thoroughly to make a 200pg/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube is used 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 | 1.84 pg/mL | |||||||||||||||||||||||||||||||||
| 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 33 (IL-33) 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 Interleukin 33 (IL-33) 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 33 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Interleukin-33 (IL-33), also known as NFHEV and DVS 27, is a 30 kDa proinflammatory protein that may also regulate gene transcription. IL-33 is constitutively expressed in smooth muscle and airway epithelial cells. Following stimulation with IL1α or IL1β, it is upregulated in arterial smooth muscle, dermal fibroblasts, and keratinocytes. IL-33 shares structural and functional features with the IL-1 family of cytokines. It binds and signals through the IL-1RL1/ST2 receptor, activating NF-kappaB and MAP kinase. IL-33 induces the production of TH2 cell-associated cytokines, including IL-4, IL-5, and IL-13, and exerts a variety of inflammatory-related biological activities. | |||||||||||||||||||||||||||||||||
| 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 | 6.25-400pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.5 ★★★★★
Based on 2395 reviews
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Product Reviews
★★★★★ 5
A lot of fun for dogs
Color: Multi Squeaker Balls, Size: Large, Style: 4-Pack, Color: Multi Squeaker Balls, Size: Large, Style: 4-Pack
My dogs love it a lot. Balls don't fall off the balcony - good size. Great sound which catches dogs attention . Colorful . Good quality.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 28, 2026
★★★★★ 5
👍🏼
Color: Multi Squeaker Balls, Size: Medium, Style: 4-Pack
This dog squeak ball has been a big hit in my home and keeps my dog entertained for long periods of time. The squeaker really grabs their attention and encourages play, tapping into their natural instincts to chase and fetch.
My dog especially loves how it bounces and squeaks, making it perfect for both indoor and outdoor play. It’s lightweight and easy for them to carry around, even for quick games of fetch in the yard.
In terms of durability, it has held up pretty well so far. The squeaker may not last forever with heavy use, but the ball itself still stays in good shape. Overall, it’s a fun, engaging toy that keeps my dog active and happy.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 29, 2026
★★★★★ 4
budget friendly for dogs who lose balls in the creek
Color: Multi Squeaker Balls, Size: Medium, Style: 4-Pack
Not great quality, but works for our needs. I have a ball crazy lab. She loves for me to throw a ball using the Chuck-it Ball Launcher. We do this in our wide back yard that butts up to a creek. At some point during her run and fetch, she will take a quick detour right into the creek. She nearly always still has the ball, but once in awhile it is lost. Not wanting to chance it on more pricy (and more durable) balls, I get these or something like these. They dont cost much, yet they do the job. I' not upset if one gets sacrificed to the creek waters and mud.
Since the balls end up getting wet often, they will kinda fall apart over time (chasing, kicking, throwing, retrieving, dog saliva etc). But she doesn't chew on them. Glad she doesnt as tennis balls arent good on teeth, but even if it was ok, the balls probably would be breached easily.
SO get these if you are like me and want a ball that does the job but arent a total financial loss if get lost or destroyed. For indoor play we and by we I mean our 3 dogs demand that we use the rubbery max glow chuck-it ball. And will ignore this style ball.
Yes, I am well trained. lol
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on November 11, 2024
★★★★★ 1
These balls SUCK! External squeakers, minimal bounce and blue that is too dark.
Color: Multi Squeaker Balls, Size: X-Small, Style: 8-Pack
We have 2 little dogs in our home - a 1.5 year old Pomsky, and a 9 year old Pekingese. My Peke has shown no interest in tennis balls for years, until today! Imagine my excitement! Until she promptly destroyed the first squeaker on one of the blue balls. When I removed it, she began peeling the logos off the ball, and then started tearing the fuzz off! My dog has never destroyed any toy in her life, so this was shocking! Then the Pomsky decided that she wanted to play fetch with the new balls - she has Golden Retriever energy - and decided to destroy a squeaker in the midst of playing! She also wasn't nearly as interested in these tennis balls as her other tennis balls that she inherited from my Peke.
1. STUPID design, with the squeakers sticking out of the balls (the don't sit flush), so they are asking to get chewed or pulled out by any dog, even elderly 10 lb dogs! And a 9 lb young adult.
2. They are difficult to squeak compared to their Kong tennis balls, hence why the Pomsky wasn't interested for long.
3. They don't bounce as much as the Kong tennis balls, either, which made them less than exciting for the athletic and agile Pomsky.
Meh. We wanted to get the smallest tennis balls we could get (1.5 inch) because neither dog will play with the 2 inch or bigger ones, but we also wanted to get colors they could easily see! The Pomsky clearly prefers the yellow balls to any other color (red, orange, green, darker blue, watermelon design), so we wanted to get like a bright blue or turquoise couple of balls to appeal to them! Well, my senior Peke liked the blue ones the most, but also liked the red and orange. Pomsky didn't like the blue ones. I think they're too dark of a blue to be easily seen in our home afternoons and evenings in the winter.
We love Outward Hound toys in our house, but these were an expensive FLOP. They are the right size, but WHY did they put the squeakers so visible and appealing to dogs? I've never seen the squeaker in the Kong tennis balls.
My advice is that Kong seems to make the best tennis balls for toy breeds, in the 1.5" size. Outward Hound dropped the ball here... no pun intended. The blue is darker than the photos depict (may be better in natural sunlight outdoors), the squeakers are external, and they barely bounce at all. Our next step will be to get the pressurized bright blue tennis balls from another brand, in hopes that the bounce and visibility will outweigh the lack of squeakiness. If those don't work, I guess we'll stick to the classic neon yellow Kong tennis balls! At my local Target, they only have multicolor packs of them, and the dark blue, red and green get completely ignored.
*Dogs can only see shades of blues, yellows and black, grey and white. While some dogs may not seem to care about color, our Pomsky clearly does. :(
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on December 21, 2023
★★★★★ 5
My Dog Loves Them
Color: Multi Squeaker Balls, Size: Small, Style: 8-Pack
These cute little squeaky balls are the perfect size for my Shih Tzu. She loves to chew on them and make them squeak all the time, so they are durable. I would purchase them again.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 16, 2026
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