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Description
Rat PAI1 ELISA KitProduct Specification Usage Required experimental equipment: 1. Microplate reader (450nm) 2. High precision pipettes and pipette tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37C incubator 4. Distilled or deionized water Sample handling and requirements: Plasma: Collect the specimen using EDTA or heparin as an anticoagulant. Centrifuge at 1000g for 15 minutes at 2 8C within 30 minutes of collection. Remove the supernatant for testing or store at
Product Specification
| Usage | Required experimental equipment: 1. Microplate reader (450nm) 2. High-precision pipettes and pipette tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37°C incubator 4. Distilled or deionized water Sample handling and requirements: Plasma: Collect the specimen using EDTA or heparin as an anticoagulant. Centrifuge at 1000×g for 15 minutes at 2-8°C within 30 minutes of collection. Remove the supernatant for testing or store at -20°C or -80°C, but avoid repeated freezing and thawing. Pre-test preparation: 1. Remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Prepare a gradient standard working solution: Add 1 mL of universal diluent to the lyophilized standard. Let stand for 15 minutes to completely dissolve, then gently mix (concentration 20 ng/mL). Then dilute to the following concentrations: 20 ng/mL, 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, and 0 ng/mL. Serial dilution method: Take seven EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 20 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 10 ng/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 uses 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 a capture antibody against Plasminogen Activator Inhibitor 1 (PAI1). 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 Plasminogen Activator Inhibitor 1 (PAI1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Plasminogen Activator Inhibitor 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Plasminogen activator inhibitor 1 (PAI-1), also known as endothelial prothrombin activator inhibitor or serum protein E1, is a protein encoded by the SERPINE1 gene. Elevated PAI-1 is a risk factor for thrombosis and atherosclerosis. PAI-1 is a serine protease inhibitor (serpin) that functions as the primary inhibitor of tissue prothrombin activator (tPA) and urokinase (uPA), the latter of which is a prothrombin activator and thus a factor in fibrinolysis (the physiological breakdown of blood clots). It is a serpin protein (SERPINE1). Another PAI, plasminogen activator inhibitor-2 (PAI-2), is secreted by the placenta and is present in significant amounts only during pregnancy. Furthermore, protease inhibitors act as inhibitors of tPA and urokinase. However, PAI-1 is the primary inhibitor of plasminogen activators. The PAI-1 gene, SERPINE1, is located on chromosome 7 (7q21.3-q22). | |||||||||||||||||||||||||||||||||
| 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 | 0.31-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | plasma |
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4.3 ★★★★★
Based on 16 reviews
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Product Reviews
★★★★★ 5
Great product and great service
Size: 16oz
By all accounts it's a great product! It arrived and I feel more confident having it with me in the car.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 28, 2026
★★★★★ 5
Essential Emergency Kit That Every Vehicle Should Have
After experiencing a flat tire on a remote highway late at night, I realized how unprepared I was for roadside emergencies. That experience led me to purchase the AUTOWN Flat Tire Repair Kit, and I'm incredibly grateful to now have this comprehensive solution in my vehicle. This kit has already saved me twice from having to call for expensive roadside assistance, and the peace of mind it provides is absolutely priceless.
The 31-piece kit is impressively complete. It includes everything you need to handle a flat tire emergency, from the tire plugs themselves to insertion tools, rasps, and even extras that many cheaper kits omit. The variety of plug sizes means you can handle different types of punctures, and having multiple plugs means you're prepared for more than one emergency before needing to restock.
What sets this kit apart is the quality of the tools. The T-handle insertion tool feels solid and well-made, not like the flimsy plastic tools that come with some budget kits. The reamer tool cuts through damaged rubber cleanly and efficiently, making it easy to prepare the puncture hole for the plug. Having used cheaper tire repair kits in the past that broke during use, I really appreciate the durability and reliability of these tools.
The storage case is another thoughtful feature that makes this kit practical for vehicle storage. Everything fits neatly in the organized case with dedicated spaces for each component, which means you can quickly find what you need during an emergency. The case is compact enough to fit in a glove box or under a seat, but it's also rugged enough to withstand being rattled around in a trunk without falling apart.
Using this kit is surprisingly straightforward, even for someone without extensive automotive experience. The process involves identifying the puncture, using the reamer to prepare the hole, inserting the plug with the T-handle tool, and trimming the excess. I was able to successfully repair a puncture on my first attempt following the basic instructions, and the repair has held perfectly for months of daily driving.
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Reviewed in the United States on February 2, 2026
★★★★★ 5
Complete and very handy tire repair kit
I bought this tire repair kit to keep in my vehicle for emergencies, and it has already come in handy. The kit comes with everything needed to fix a flat quickly, and having all the tools in one case is very convenient.
The plugs worked well and sealed the puncture properly, allowing me to get back on the road without issues. The tools feel solid and easy to use, even if you don’t have a lot of experience with tire repairs.
The case is a bit tight once everything is packed back in, but that’s a small issue. Overall, this is a reliable and practical kit that’s great to have for cars, motorcycles, or other vehicles. I’m very glad I bought it.
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Reviewed in the United States on February 2, 2026
★★★★★ 5
Great in every way
Nice kit full of useful tools to fix punctures in tires better than anything I have ever tried before. The quality is pretty good - good enough to do what it’s supposed to do. It’s better if you fix your tire when it’s off of your car. It’s a lot easier to push the plug down when your standing and can use your body weight rather than laying on the ground in an awkward position and trying to push the plug in. I did the latter and it was not fun. I lived and learned. Anyhow, I wasn’t going to drive on rims essentially with a flat to get it repaired somewhere so this was a fantastic buy.
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Reviewed in the United States on May 17, 2026
★★★★★ 4
Nice kit
Nice kit, works well
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Reviewed in the United States on May 18, 2026