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Description
Mouse IL1RA 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. Other biological fluids: Centrifuge at 1000xg for 20 minutes, remove the supernatant, and test. 5. Pre-test preparation: 1. Remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Preparation of 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 2000pg/mL). Then dilute to the following concentrations: 2000pg/mL, 1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.25pg/mL, and 0pg/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each tube. Pipette 500uL of the 2000pg/mL standard working solution into the first EP tube and mix thoroughly to make a 1000pg/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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| Theory | This kit utilizes a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA). Sample, standard, biotinylated detection antibody, and HRP conjugate are sequentially added to microwells pre-coated with an Interleukin 1 Receptor Antagonist (IL1RA) 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 1 Receptor Antagonist (IL1RA) 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 1 Receptor Antagonist ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Interleukin-1 receptor antagonist (IL-1Ra) is a protein encoded by the IL1RN gene. Originally known as an IL-1 inhibitor, it is an agent that nonfunctionally binds to the interleukin-1 receptor (IL-1R) on the cell surface, the same receptor to which interleukin-1 (IL-1) binds, preventing IL-1 from signaling to that cell. A member of the interleukin-1 family of cytokines, IL-1Ra is secreted by various cell types, including immune cells, epithelial cells, and adipocytes, and serves as a natural inhibitor of the proinflammatory effects of IL-1β. This protein inhibits the activity of interleukin-1 alpha (IL1A) and interleukin-1 beta (IL1B), modulating various interleukin-1-related immune and inflammatory responses. | |||||||||||||||||||||||||||||||||
| 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 | 31.25-2000 pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenates and other biological fluids |
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4.4 ★★★★★
Based on 24 reviews
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Product Reviews
★★★★★ 5
Toddler loofah set worth the buy !
Color: Frog&strawberry
The cutest loofah set I could find! So soft, thick, and my kids love the theme. I love how the bundle is, it’s perfect for my son and daughter. The sponges are not rough on their skin and are very durable!
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Reviewed in the United States on May 13, 2026
★★★★★ 4
Suave al contacto para restregarle el cuerpo a mi nene
Color: Turtle&bunny
Me a encantado este producto lo único malo que me pasó es que la parte de la esponja se descoció o me tocó que cocerlo para que no se le saliera la esponja, de ahí en fuera muy buen producto
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Reviewed in the United States on March 23, 2026
★★★★★ 5
Made very well!~!
Color: Frog&strawberry, Color: Frog&strawberry
Arrived as pictured. Boxed nice and cute. Suitable to send as a gift or for retail sales. These are not only adorable but wash very well. My grandson and I both like them. He said the strawberry was for me and he took the frog. I normally have to bribe him to bathe but all I have to say is froggy is waiting and he runs in!~!~ I love my strawberry and it really cleans well. Such fun and I'm delighted with them both. The size is perfect and I'm grateful. We have been using them for awhile to and they have held up like new. Worth the cost and priced very fair.
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Reviewed in the United States on May 3, 2026
★★★★★ 5
Super Cute Sponges for kids!
Color: Frog&strawberry, Color: Frog&strawberry
This is a pack of super cute kids bath sponges. These are sponges attached to loofahs that have a loop on them and can be hung to dry. These are shaped, One looked like a frog and the other like a strawberry making this pack accommodate both boy and girls. These sponges work very well and feel like they're made of a decent quality. The sponge area is very soft and the loofahs feel well made. These are a perfect size for the kiddos. These do a good job at lathering up the suds for the little ones. These also feel like they have a good durability and don't feel like they're gonna fall apart anytime soon. These are super cute and would make a great gift as well.
The value on this is not very expensive. These are affordable especially for gifting. giving this an expected value in my opinion and at the time of this review.
Overall, these sponges are super cute and would make a fantastic gifts for little ones. I would have no issues recommending these.
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Reviewed in the United States on March 31, 2026
★★★★★ 5
Very functional, and lots of fun for little kids. Gentle enough for their skin, too.
Color: Frog&strawberry
These are so awesome. Two of them come in a box. There's a frog one and a strawberry one. They are inside individual zip lock bags inside the box. There's a scrubbie/loofa (not a natural one, just the plastic webbing type) on one side, and cloth-covered plushie/sponge on the other side that is the strawberry/frog. My granddaughter loves them. She can use them to clean and scrub herself, just like a big girl, but with a fun sponge/scrubby that she likes, so she always knows that there's a special one that is just hers. They absorb plenty of water and soap, and are good for cleaning their whole body. They are just the right size to fit in an adult hand, so a little big for solo use by a little hand, but there are plenty of bits to grab onto on either side, so it's not hard for a little kid to hang onto them. The loofah/scrubby side is well made, although it will become big and loose as those things always do if you pull on them too much. The sponge/toy side is very soft, and definitely gentle enough for a small child's skin.
My only dislike is the small loop for hanging it on a hook in the shower. The little loop is just tiny and hard to separate, so it's difficult to get it onto a hook so it can hang and dry out in between uses. It would have been very easy to make the loop longer and more functional, so I'm not sure why they didn't.
Other than that one, minor inconvenience, this is a great product.
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Reviewed in the United States on February 26, 2026