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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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4.1 ★★★★★
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Product Reviews
★★★★★ 5
Clean Look, Super Comfortable – Instantly a Favorite
Size: 9.5, Color: White Leather
These sneakers are exactly what I was looking for—sleek, minimal, and incredibly comfortable right out of the box. The white leather looks sharp and pairs easily with everything from jeans to chinos, even dressed-down office wear.
What surprised me most was the comfort. The sole is lightweight with just the right amount of cushioning, making them perfect for walking around all day. I’ve worn them on long city days and never felt sore or tired.
They fit true to size (I’m a 9.5 and they’re spot on), and the quality feels premium. If you're after a versatile white sneaker that balances style and comfort, these are 100% worth it.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 29, 2025
★★★★★ 5
Comfortable
Size: 10.5 Wide, Color: Black Leather/Black
Very comfortable shoes, you feel like walking on clouds
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Reviewed in the United States on April 26, 2026
★★★★★ 3
Not for all surfaces.
Size: 11.5 Wide, Color: Black Leather/Optic White
The sneakers look great and are comfortable on flat, smooth surfaces. Warning: The sole is very flexible and not well cushioned so a walk on rough or uneven surfaces can become uncomfortable quickly. Just understand the limitations.
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Reviewed in the United States on June 1, 2026
★★★★★ 5
⭐⭐⭐⭐⭐ Clean Look, Comfortable Ride
Size: 11.5 Wide, Color: Woodbury Handstain
The Cole Haan Men’s GrandPro Tennis sneakers are a perfect mix of laid-back and polished — the kind of shoe you can wear everywhere without thinking too hard about it.
Right off the bat, the design is subtle but refined. They’re minimalist enough to pair with jeans and a hoodie, but they also work with chinos and a button-down for a more put-together look. The “tennis” aesthetic keeps it classic without feeling like you’re wearing athletic shoes.
Comfort is where these really shine. They’re lightweight and cushioned in all the right places, so long walks, errands, or standing around at events don’t end with sore feet. There’s no stiff break-in period — they feel good from the first wear. The sole has just the right balance of support and flexibility, which makes them easy to wear all day.
The materials feel good too — soft, sturdy, and not flimsy. They hold up well after repeated wear and still look fresh even after a few outings. They don’t crease or scuff up excessively, which is always nice.
If I had to point out a downside, it’s that they’re not performance sneakers for athletics — but that’s not what they’re meant to be. They’re lifestyle shoes that look great and feel great for everyday wear.
Overall, a solid go-to sneaker that elevates casual outfits and keeps comfort front and center. Easy recommend if you want versatile kicks that stay stylish without being loud.
I was able to pick up this particular pair here on Amazon for 50 bucks, 56 total with everything and honestly I’m still scratching my head in that deal. I mean my New Balance were 70 bucks or something like that.
These are brown leather, sophisticated and comfortable, just amazing shoes. I get compliments all the time.
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Reviewed in the United States on February 16, 2026
★★★★★ 5
Quality and Comfort
Size: 12, Color: White
My husband is a Cole Haan shoe person and what we love mostly is the comfort and quality and style.
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Reviewed in the United States on May 13, 2026