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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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★★★★★ 3
Cheesy romance that hits you in all the feels.
A cheesy romance full of tropes but it pulls them off and keeps you smiling and rooting for the main characters the whole way through.
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Reviewed in the United States on December 17, 2019
★★★★★ 5
Entertaining
Format: Paperback
This took me four days to finish while taking classes over all good though kind of predictable
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Reviewed in the United States on May 26, 2026
★★★★★ 4
Great enemies to lovers
Great tension, love the banter, wish it had more kiss scenes and description when they did BUT it was amazing! Read it in one day. Fun read, do recommend!!!!
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Reviewed in the United States on March 11, 2025
★★★★★ 5
The best book I have ever read
The best book I have ever read, I would read it 100 times more. I recomend it a ton. Go buy it now!
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Reviewed in the United States on April 18, 2026
★★★★★ 5
The hate you give; that end up making me LOVE YOU
You can tell that Ally had so many doubts when it came to Chase... Especially when it came to her feelings for him and how he felt for her. I figured he had a thing for her even though what he did at the dance freshman year was a bust... though it was the right thing to do for Ally though I believe he should've given Ally an option to decide that decision with Declan... her doubts with Chase has the present intervening with the past which had proven to see him being so different... but things can change....I think he hated her ONLY because she hated him... & jumped to a conclusion before he could explain to her why he did what he did at the dance... Same when she accused him of the photo when she went to that club and dancing on the pole.. she accused him and came to a conclusion that he did it... she only found out that he didn't do it... because Declan and Chase got into it; by Chase demanding him to apologize... Declan wasn't even worth the air she breathes in senior year or freshman year if you look at it.... plus Chase parents are horrible especially his father sheesh no one wants a parent like that...
But oh.... when he (Chase) said: "I didn't have a choice, Ally, because if I didn't hate you, I would love you, and loving you would only break my heart." I knew that had to be the reason, and I knew he liked/loved her, but she was to blind to see through her hate for him... by getting mad about Declan who was the same person he was freshman year to senior year. I figure I was right too... but kind sad and thinking duh Ally when she stated: "I'd created an enemy out of a boy whose only fault was loving me, and my heart was breaking from his words." Sometimes you learn from your mistakes... but half the time I thought Tessa set Ally up with Chase so that feelings can start to show on Ally side since Chase already had feelings. No way; you think you not like someone; can't get rid of someone if you "hate them".. when you can you actually can't stand them; when you don't think you have feelings. No way I will be around someone or try rid them if I dislike them for bullying me down or hurting my feelings...they wouldn't be in the same home heck I wouldn't care what my daddy said... build me a treehouse I don't have time for living in a house with the enemy... in reality Chase was not Ally enemy he was her lover she just didn't know it yet.... but at least, in the end, he showed her that no matter what I'm coming for you "to make you my girl; because I always love you" and she finally stops running and agree to date him once she found out that her father knew about Chase loving her among other things. But besides that... UGH! I need a book with Ally twin brother Shane and her best friend, Tessa... I would like to see them together because she has a thing for him and he doesn't even know!!! But maybe he does!
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Reviewed in the United States on June 28, 2019