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Description
Mouse PAI1 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: 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: 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. Plasma: Collect specimens using EDTA or heparin as anticoagulants and centrifuge them at 1000×g for 15 minutes at 2-8℃ within 30 minutes of collection. The supernatant can be tested or stored at -20℃ or -80℃, but repeated freezing and thawing should be avoided. Other biological fluids: Centrifuge at 1000xg for 20 minutes, remove the supernatant, and test. Pre-test preparation: 1. Remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Prepare the standard gradient working solution: Add 1 mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 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 7 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 directly as a blank well; there is no need to aspirate the liquid from the penultimate tube. See the figure below for details. 3. Preparation of Biotinylated Antibody Working Solution: 15 minutes before use, centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration using universal diluent (e.g., 10µL concentrate + 990µL universal diluent). Prepare immediately before use. 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 | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Plasminogen Activator Inhibitor 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | PAI-1, also known as endothelial prothrombin activator inhibitor or serum protein E1, is a serine protease inhibitor (serpin). Its primary function is to inhibit urokinase plasmakinase activator (uPA), an enzyme responsible for cleaving plasmakinogen to form plasmakin. Plasmin, either by itself or in conjunction with matrix metalloproteinases, mediates the degradation of the extracellular matrix. In this context, PAI-1 inhibits uPA by binding to its active site, preventing plasmakin formation. Additional inhibition occurs through PAI-1 binding to the uPA/uPA receptor complex, leading to its degradation. Therefore, PAI inhibits the serine proteases tPA and uPA/urokinase, thereby acting as an inhibitor of fibrinolysis, the physiological process that degrades blood clots. Furthermore, PAI-1 inhibits the activity of matrix metalloproteinases, which play a key role in the invasion of malignant cells through the basal lamina. | |||||||||||||||||||||||||||||||||
| 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 | Serum, plasma, and other biological fluids |
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4.1 ★★★★★
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★★★★★ 5
Agile Practice Guide
Format: Paperback
Una muy buena compra para tener como guia repaso de los conceptos mas importantes de las metodologías agiles, y que variables tener en cuenta a la hora de encarar un proyecto y definir con que metodología se gestionará el mismo.
En un mundo en donde todos quieren IMPONER metodologías que todavía no entienden muy bien pero estan de moda, es una muy buena herramienta para entender cuando es valido hablar de metodologías agiles, de cuando hablamos de metodologías hibridas, o inclusive discutir si no es un approach mas eficiente el de ejecutar una metodología waterfall o lineal.
100% recomendado.
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Reviewed in the United States on April 23, 2024
★★★★★ 5
Excellent Introduction to SQL
Format: Paperback
I am compelled to share my overwhelmingly positive experience with the SQL Quickstart Giude which has proven to be an exceptional resource for anyone wanting to delve into the world of SQL coding.
First and foremost, the book is written in an extremely accessible manner, deftly simplifying complex concepts into digestible portions for beginners, yet providing deeper insights for seasoned developers. It transcends the barrier of technical jargon, presenting information in an intuitive, reader-friendly style.
However, the true selling point of this book, which separates it from the plethora of SQL resources available in the market, is the ingenious inclusion of its online resources. This addition enhances the book's value immeasurably, allowing the reader to step beyond the theoretical and put their knowledge into practice immediately.
The online resources that accompany the book provide a unique learning ecosystem, an ideal blend of theory and practice. They offer a plethora of well-organized and insightful coding examples for each chapter, which are also available for download. This allows the readers not only to work along as they learn but also to experiment and implement their own iterations of the code. This hands-on approach cements the concepts learned in a way that theoretical learning alone cannot.
Moreover, these downloads enable offline access, which ensures that you can learn at your convenience, a feature that not many SQL books offer. This aspect of anytime, anywhere learning is crucial in our current fast-paced world.
Furthermore, the author provides ample exercises to test your understanding, which are then complemented by detailed solutions on the online platform. This real-time feedback is instrumental in understanding one's progress and identifying areas for improvement, which paves the way for a confident and competent SQL programmer.
In conclusion, the "SQL Quickstart Guide" with its companion online resources has proven to be an unbeatable combination for learning SQL. It has created a learning experience that is seamless, engaging, and, most importantly, effective. It is a rare gem in a field often dominated by dense, inaccessible texts and I would highly recommend it to anyone wishing to embark or advance their journey in SQL programming.
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Reviewed in the United States on July 24, 2023
★★★★★ 5
Easy to follow, well written book. Can easily be used in liue of a textbook. Get spiral edition.
Format: Spiral-bound
Walter Shields does a fantastic job of writing this book from a perspective that a reader will have no clue on databases, or SQL. I'm a former Database programmer, but after being away for so many years I had to come back to SQL, and I needed a reference quick. Everything is written in a step by step easy to swallow informational way that leaves no room for failure. Even the examples are fantastic. This book is compatible with opensource SQL so need to pay for anything but the book.
Get the spiral edition, it lays flat on the table and is easier to follow.
Unlike McGraw Hill, Cengage, Thomson Course Technology, this book doesn't expire and block you from buying a used copy to prevent you from downloading the files needed to learn.
A fantastic reference, a great book, timeless because it doesn't expire.
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Reviewed in the United States on October 22, 2024
★★★★★ 4
SQL qsg book
Format: Paperback, Format: Paperback
I've taken a couple classes on SQL some time ago and was looking for a book to go over basics. This is a perfect book to start with the basics of understanding how to write a SQL query. I really liked that there is a free database to use in SQLite to follow along with the examples in the book. That was very helpful for me. I appreciated the tips throughout the chapters and the practice questions at the end. Easy to read and follow.
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Reviewed in the United States on April 27, 2023
★★★★★ 5
This is a good very distant overview
Format: Paperback
Sometimes I need a really simplistic overview of an entire subject before I feel comfortable sitting down to learn it. Like if you are going to learn about, say, 13th Century France you might want to know just a little about the major issues of the entire century before sitting down with a text book. This book is good for that. It's short, it gets you started with the main keywords, and at the end you have a good picture of what the language can do. And you can get to the end in a day or two. It's very well written, well organized, clear, and very very limited in scope. But if you read it, you will be in good shape to start getting down to business with working with SQL with a more advanced text. This book, in my opinion, was well worth the money and the read even though ultimately I turned to something far more advanced.
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Reviewed in the United States on October 12, 2024
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