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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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4.2 ★★★★★
Based on 2316 reviews
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Product Reviews
★★★★★ 5
Colorful and handy
These are really nice and also inexpensive. The colors are bright and similar to photo. The wrist strap is handy. They fit the appletv control just right. (Snug enough but not hard to take on and off). These help protect against falls and also make it easier to find the controls. No complaints
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Reviewed in the United States on October 23, 2019
★★★★★ 5
Excellent quality. Highly recommended!
Excellent quality and packaging was very secure and minimal but recyclable! I love the cell of the remote control of silicone skin. Both a nicely vibrant colors and easy to find. The worst straps are seriously heavy duty. That's great for gaming. I didn't need them but the silicone protection is amazing. I have two Apple TVs. The price you can't beat anywhere else.
If you've ever dropped an apple remote, you quickly discover that the remote is partially made of glass. They are expensive to replace.
Save yourself a headache and get these.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 28, 2021
★★★★★ 5
Prepare to love your Apple remote again!
This is a must buy for anyone that owns an Appte TV. The remotes are small, and sleek, and slippery. My whole family drops or loses the remote constantly. It was a constant struggle.
After slipping on the teal blue silicon sleeve it was like we had a whole new remote.
It makes the remote much less slick feeling. Not nearly as easy to drop.
The bright blue color makes it super simple to spot in the couch cushions or on the counter.
The sleeve provides protection from the all too common drops that happen everyday.
It does make the remote slightly thicker, but that isn't a bad thing. It fits in your hand much better and makes operation easier too.
Well worth the price and you get two!
Totally recommend these.
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Reviewed in the United States on December 5, 2019
★★★★★ 5
Great value, solves a big problem!
Wow, what a great product, so inexpensive for two, and solved one of my biggest aggravations. This remote fits perfectly over the apple TV remote. The remote is so slippery, black, and it is not designed to be able to tell which way is up. This cover solves every problem. It is no non-stick, I don't lose it anymore. I used to drop it once a day at least - no more dropping the remote. It fits just right, you can easily tell which way is the up. It fits just right, and you can still use every feature of the touch pad, the fast forward, etc. Just right!!! Excellent!!!
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Reviewed in the United States on October 10, 2019
★★★★★ 4
Great fit, tether / lanyard probably won't stay on long
A great fit and deal for the money. It's well-balanced in the hand, and makes the remote much less awkward to use. I have two Apple TV's in the house, and the different colors help keep the remotes straight.
There's a lanyard that attaches to a small little bit of silicone. Having the lanyard is nice, but it doesn't stay connected because the bit of plastic is so small, it rips even under a small force. It's still completely useful and worth the purchase to me without the lanyard, but if the lanyard is a dealbreaker for you, you may wish to reconsider. I can't imagine it staying on very long in any environment. 2 kids and a cat it lasted about 20 minutes.
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Reviewed in the United States on December 4, 2019
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