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Description
Rat IL33 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 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 Interleukin 33 (IL33) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase and to yellow by acid. The intensity of the color is positively correlated with the amount of Interleukin 33 (IL33) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat 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 | 15.6-1000 pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.6 ★★★★★
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Product Reviews
★★★★★ 4
A little drying, but effective at reducing white heads and build up -- will keep using!
Size: 8 Fl Oz (Pack of 1)
I have very delicate skin that is prone to acne and eczema. I also have a lot of allergies that prevent me from using most skin products so I was pretty nervous about trying a new one, but I gave it a chance.
The first couple of days I noticed some itchiness in my skin, but it was effectively clearing dead skin and reducing whiteheads so I kept using it and the itching went away.
My skin does feel a bit dry after using it, and it probably would be better alongside the vanicream moisturizer; however, I try to only start new products one at a time so that I can easily pin point things that may be bothering me.
As far as pricing, I think it's pretty affordable considering it comes in a fairly large bottle and you only have to use a pea-sized amount each wash.
It also doesn't leave any weird films or grease, and doesn't seem to have any weird smells.
Altogether, really like the product over the week or two that I have been using it, and my skin looks a lot cleaner and more evenly textured.
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Reviewed in the United States on May 16, 2026
★★★★★ 5
Best cleanser I’ve ever had!
Size: 2.5 Fl Oz (Pack of 1)
This is absolutely the best cleanser I’ve ever had. My skin is dry and sensitive, and very reactive, so I’ve always had a hard time finding good products. But this cleanser is honestly the best. It’s never broken me out, doesn’t dry out my skin, and leaves it feeling so soft. It isn’t greasy at all, and it’s completely unscented, which I love. Super effective and lightweight, washes off easily, and good enough to use day and night if you want. I use this daily, and it’s honestly great. Will be repurchasing once I’m done mine!
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Reviewed in the United States on April 23, 2026
★★★★★ 5
Works Great, Fragrance-Free & Lathers Well
Size: 2.5 Fl Oz (Pack of 1)
I recently became highly allergic to all artificial fragrances and beeswax so this Sensitive Skin Vanicream Gentle Facial Cleanser was recommended by my dermatologist. I tried a few other brands, and this one is my favorite by far. It lathers well, is not greasy, and leaves my skin feeling soft and clean. I like to use the 8 oz bottle size with the pump at my sink and in my shower, but this 2.5 oz size was perfect for 2 weeks in Europe, and I still have a lot left with using it twice a day. Also, it doesn't burn my eyes, so I would say it is safe for taking off eye make up. Highly recommend!
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Reviewed in the United States on May 27, 2026
★★★★★ 5
A total lifesaver for seb derm!
Size: 8 Fl Oz (Pack of 1)
I honestly don’t know what my husband would do without this cleanser. He struggles with seborrheic dermatitis, and finding something that doesn’t trigger a flare-up is usually a nightmare, but this stuff is absolute gold. It’s so gentle on his skin, no fragrance or weird oily texture and the fact that it’s less than $10 is honestly a steal for the amount you get in a bottle.
We’ve repurchased multiple bottles at this point because it’s a total staple in his routine now. He has tried EVERYTHING even prescribed stuff from the dermatologist but nothing worked until we found this.
It’s also super easy to dispense and use with the pump, so there’s no mess or hassle. If you're looking for something that actually works for sensitive skin without breaking the bank, just get it!
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Reviewed in the United States on March 28, 2026
★★★★★ 5
The pads have changed but still a good product
I have been using these make up remover pads for many years now and will continue to use them. I like the 3 pack so that I do not run out. I usually only use 1 pad a day so a 3pk will last me for a long while. I do not have any issues with the pads drying out in the time frame from opening them to using the last one so that is a major plus. I have no issues with these on my skin or that close to my eyes and they do a great job of removing my mascara, which is the only make up I use. I recommend them. I will say that in 2025 they have changed the material for their pads, they are now thinner and do not have the texture they used to have which for me the only difference is that they can be a bit more difficult to separate but nothing to complain about.
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Reviewed in the United States on November 7, 2025