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Description
Rat IL25 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 25 (IL25) 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 25 (IL25) 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 25 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | IL-25 is a cytokine that belongs to the IL-17 cytokine family, along with IL-17A (also known as IL-17), IL-17B, IL-17C, IL-17D, and IL-17F. All members share homologous amino acid sequence segments and spatially conserved cysteines. However, it is IL-25 that differs from the other members in terms of function and structure. IL-25 has a heterodimeric receptor composed of two subunits, IL-17RA and IL-17RB. The IL-17RA subunit is also common to IL-17A and IL-17F, while IL-17RB is also shared by IL-17B. Both subunits of the receptor are essential for IL-25's function. IL-25 is produced by many cell types, including T cells, dendritic cells, macrophages, mast cells, basophils, eosinophils, epithelial cells, and Paneth 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 | 7.8-500 pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.3 ★★★★★
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Product Reviews
★★★★★ 5
Durable, Odorless, and Reliable — A Must‑Have 3‑Pack
Size: 3Pack 16"x20" Black
I’m really impressed with this 3 Pack PTFE Teflon Sheet for Heat Press (16 x 20"). The quality is excellent, and the non‑stick surface makes every press come out clean and smooth. One thing I really appreciate is that there’s absolutely no smell, even when using high heat. It makes the whole process more comfortable and professional.
These sheets offer great protection for both the heat press and the materials. They prevent scorching, keep the plates clean, and stop any vinyl or ink from sticking. My projects come out more consistent because the sheets help distribute heat evenly and act as a reliable barrier.
The 3‑pack is a huge bonus. I keep one on my press, one as a backup, and one for different materials like HTV or sublimation. It saves time and keeps everything cleaner when I’m working on multiple items.
The 16 x 20 size is perfect for shirts, tote bags, and larger designs, and the sheets stay flat without curling. If you want something that protects your equipment, has no smell, and delivers clean results every time, this set is definitely worth it.
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Reviewed in the United States on December 18, 2025
★★★★★ 5
T shirt Quilt Use
Size: 3Pack 16"x20" Black
I bought this specifically for ironing interfacing onto t-shirts before cutting into squares. Some of the memory t-shirt's had ‘puffy paint’ and other shirts had uncertain designs that may be ruined by ironing without protection or by using just a pressing cloth (cotton material). A teflon sheet was suggested on a you tube video to ‘play it safe’ when making a memory quilt. I felt the heat resistance teflon helped because you need to hold iron in same place for a longer time than ‘regular ironing’ movement. It worked well, the only (minor) issue is you can’t see through it. It did not interfere at all because positioning of shirt and interfacing was in place. I’m a ‘newby’ at this process; this worked for me. This is a good quality material; one sheet can be used for ironing t-shirts forever, I plan on using the other two teflon sheets to line outdoor BBQ drawer.
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Reviewed in the United States on April 29, 2026
★★★★★ 5
High Quality & a Must-Have for Any Heat Press User
Size: 10Pack 12"x16" Black
These PTFE sheets are an absolute must if you use a heat press. I use them all the time and the quality is excellent. They’re thick, durable, and truly non-stick, which makes pressing vinyl so much easier and cleaner.
They protect my projects and my heat press perfectly, and they hold up well even with frequent use. I also love the size—plenty of coverage for different projects. This 10-pack is a great value for the price, especially if you’re pressing often like I am. I’ll definitely be keeping these on hand and repurchasing when needed.
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Reviewed in the United States on January 24, 2026
★★★★★ 5
Must have
Size: 3Pack 16"x20" Black
These Teflon sheets are a must-have for heat press projects! They protect my designs and heat press perfectly without sticking or leaving any marks. The material feels durable, reusable, and easy to clean after multiple uses. They handle high heat really well and make crafting so much smoother and less stressful. Perfect for sublimation, HTV, and other transfer projects. Great quality and definitely worth buying for anyone who does crafting or custom apparel!
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Reviewed in the United States on May 12, 2026
★★★★★ 4
They do the job
Size: 3Pack 16"x20" Black
They work well.
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Reviewed in the United States on June 9, 2026