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Description
Rat PAI-2 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 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 100 ng/mL). Then dilute to the following concentrations: 100 ng/mL, 50 ng/mL, 25 ng/mL, 12.5 ng/mL, 6.25 ng/mL, 3.125 ng/mL, 1.5625 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 100 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 50 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 2 (PAI-2). 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 2 (PAI-2) 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 2 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Plasminogen activator inhibitor 2 (Placental PAI, SerpinB2, PAI-2), a serine protease inhibitor of the Serpin superfamily, is a coagulation factor that inactivates tissue prothrombin activator and urokinase. It is present in most cells, particularly monocytes and macrophages. PAI-2 exists in two forms: a 60 kDa extracellular glycosylated form and a 43 kDa intracellular form. | |||||||||||||||||||||||||||||||||
| 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 | 1.56-100 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | plasma |
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Product Reviews
★★★★★ 4
Too small
Size: One Size, Color: Light Blue Shark Waves
Its too small for my kindergartener.
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Reviewed in the United States on December 9, 2025
★★★★★ 5
Pretty in Pink and Packed with Fun
Size: One Size, Color: Lilac Ditsy Floral
This cute pink floral backpack has quickly become the star of our morning routine! The bright flowers and cheerful pink color make it feel extra special—my little one beams every time she puts it on. It’s the perfect size for kids: roomy enough to fit snacks, a water bottle, and school essentials, but not so big that it looks like the backpack is wearing them.
The straps are comfy and adjustable, and the zipper is smooth (no stuck mornings here, thank goodness!). Plus, it’s lightweight, which makes it great for school, playdates, or even little adventures to the park.
It’s not just a backpack—it’s basically a portable burst of sunshine and style. 🌸💕 Definitely a winner for any kiddo who loves to carry their world in something as cute as they are!
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Reviewed in the United States on August 26, 2025
★★★★★ 5
Great quality.
Size: One Size, Color: Lilac Ditsy Floral
Love this. Perfect size for my 3yo. So cute and good quality.
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Reviewed in the United States on November 12, 2025
★★★★★ 5
Perfect toddler diaper bag
Size: One Size, Color: Black
I am a diaper bag minimalist but sometimes you just need a little extra space. This small backpack is perfect for all the essentials and more. It can easily fit a few diapers, wipes, toys and snacks along with much more. The side pockets are also able to hold things.
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Reviewed in the United States on November 26, 2025
★★★★★ 5
Perfect Toddler Backpack - Spidey Design and Great as a Diaper Bag
Color: Marvel: Spidey Kid, Style: Kids Medium
This backpack has been such a great find for us, especially with our Spidey-Loving little one. The Spider-Man design is adorable - bright, bold, and instantly loved by the little one. It's the perfect size for toddlers, but still roomy enough that we've been using it as a diaper bag, and it works surprisingly well.
The compartments are thoughtfully laid out, making it easy to organize diapers, wipes, snacks, a change of clothes, and even a small toy or two. The zippers are smooth, the material is durable, and the straps are comfortable for either of us or the little one to carry.
It's lightweight but sturdy, and the quality feels great for everyday use - whether we're heading to daycare, the park, or just running errands.
If you're looking for a toddler backpack that's cute, functional, and versatile enough to double as a diaper bag, this one is an awesome choice.
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Reviewed in the United States on April 17, 2026