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Description
Mouse IL17A 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, and remove the supernatant for testing. (10ul sample load: Serum: Place the whole blood sample collected in the serum separator tube at room temperature for 2 hours or at 4℃ overnight, then centrifuge at 1000×g for 20 minutes. Remove the supernatant, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. Plasma: Collect the specimen using EDTA or heparin as an anticoagulant, and centrifuge at 1000×g for 15 minutes at 2-8℃ within 30 minutes of collection. Remove the supernatant for testing, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing.) Preparation before testing: 1. Please remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Prepare the gradient working solution of the standard: Add 1 mL of universal diluent to the lyophilized standard. Let stand for 15 minutes to completely dissolve, then gently mix (concentration is 300 pg/mL). Then dilute to the following concentrations: 300 pg/mL, 150 pg/mL, 75 pg/mL, 37.5 pg/mL, 18.75 pg/mL, 9.375 pg/mL, 4.6875 pg/mL, and 0 pg/mL. Serial dilution method: Take seven EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 300 pg/mL standard working solution into the first EP tube and mix thoroughly to make a 150 pg/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 17 A (IL17A) 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 17 A (IL17A) 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 17 A ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | IL-17A is a protein encoded by the IL17A gene. In rodents, IL-17A was once called CTLA8 because of its similarity to a viral gene (O40633). It is a proinflammatory cytokine produced by activated T cells. This cytokine regulates the activity of NF-kappaB and mitogen-activated protein kinases. It stimulates the expression of IL-6 and cyclooxygenase-2 (PTGS2/COX-2) and enhances nitric oxide (NO) production. Elevated levels of IL-17A are associated with several chronic inflammatory diseases, including rheumatoid arthritis, psoriasis, and multiple sclerosis. | |||||||||||||||||||||||||||||||||
| 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 | 4.68-300 pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids (10ul loading serum, plasma) |
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4.5 ★★★★★
Based on 21 reviews
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Product Reviews
★★★★★ 5
Endless Fun and Excitement: Outward Hound Squeaker Ballz Fetch Dog Toy Review
Color: Multi Squeaker Balls, Size: Medium, Style: 4-Pack
f you're searching for the ultimate entertainment solution for your furry friend, look no further than the Outward Hound Squeaker Ballz Fetch Dog Toy. As a proud dog owner, I can attest to the sheer joy and excitement that these toys bring to our canine companions. Here's why the Outward Hound Squeaker Ballz Fetch Dog Toy deserves all the praise:
Irresistible Squeaky Fun: The Outward Hound Squeaker Ballz are an absolute delight for dogs of all shapes and sizes. Each ball features an attention-grabbing squeaker that ignites your pup's natural instincts for play and exploration. The squeaky sound adds an extra layer of excitement to fetch games, encouraging hours of active and engaging playtime.
Durable Construction: Made from high-quality, durable materials, the Outward Hound Squeaker Ballz are built to withstand the rigors of enthusiastic play. Whether your dog loves to chase, fetch, or chew, these toys hold up remarkably well against rough handling and gnawing, ensuring long-lasting enjoyment and entertainment for your furry friend.
Versatile Play Options: With a convenient pack of four medium-sized balls, the Outward Hound Squeaker Ballz offer endless opportunities for interactive play and bonding with your dog. Whether you're playing indoors or outdoors, these versatile toys provide a fun and engaging outlet for your dog's energy, helping to promote physical activity and mental stimulation.
Bright and Colorful Design: The vibrant colors and eye-catching design of the Outward Hound Squeaker Ballz make them easy to spot and retrieve during outdoor play sessions. Their bright hues add an element of excitement to fetch games, keeping your dog entertained and focused on the task at hand.
Safe and Non-Toxic: As responsible pet owners, we prioritize the safety and well-being of our furry companions. The Outward Hound Squeaker Ballz are crafted from pet-safe materials that are free from harmful chemicals and toxins, providing peace of mind knowing that your dog is playing with a safe and non-toxic toy.
Affordable and Value-Packed: With four balls included in each pack, the Outward Hound Squeaker Ballz offer exceptional value for the price. You can keep multiple balls on hand for backup or share them with fellow dog owners, spreading the joy and excitement of playtime to canine friends everywhere.
In conclusion, the Outward Hound Squeaker Ballz Fetch Dog Toy is a must-have addition to any dog owner's collection of pet supplies. With their irresistible squeaky fun, durable construction, versatile play options, bright and colorful design, safety features, and unbeatable value, these toys bring endless joy and entertainment to dogs of all ages and breeds. Treat your furry friend to the ultimate playtime experience with Outward Hound Squeaker Ballz Fetch Dog Toy - you won't be disappointed!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 27, 2024
★★★★★ 5
Squeaky fun!!
Color: Multi-colored, Size: Small - 12 Pack
These are perfect they fit in my hyper pet shooter and they squeak too! My dogs absolutely love themZ. Seems to be good value and I think they will soften up over time.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 28, 2026
★★★★★ 5
Small and squeaky!
Pretty perfect! Great size for small dog and they do squeak. It’s helpful to use both hands when getting the ball to squeak. It’s a bit difficult with just one hand, but that can’t be avoided because the size of the tennis ball is small. We use these inside and outside to exercise our puppy.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on November 13, 2025
★★★★★ 5
Milo approved!!!
Color: Multi-colored, Size: Large - 4 Pack
My dog, Milo, loves these squeeky balls. He is a larger dog but not huge. He's about 60 lbs. They are the perfect fit to his mouth. Soft enough for him to chew on but sturdy enough to hold up.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 30, 2026
★★★★★ 5
Squeaky Tennis Balls Are Great Fun!
Color: Multi-colored, Size: Large - 4 Pack
My Doberman loves these! That said, she doe's tear them up over time because once she gets one, she squeak's it continually and chews it. They are tennis ball which are not made to withstand aggressive chewers, but she loves them. They are a great size for her, we play with them every night. She's not a picker and doesn't swallow things so I let her have each one until it's destroyed. Then we get out another one to start the process again. They are a reasonable price for the fun she has.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 31, 2026