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Description
Rat AQP-4 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 preparation and requirements: 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 and
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 preparation and requirements: 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 and mince the tissue. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1g of tissue sample to 9mL 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 collect the supernatant for analysis. Cell Lysis Buffer: Gently wash adherent cells with pre-chilled PBS, then trypsinize and collect the cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash the collected cells three times with pre-chilled 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 freezing and thawing or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and collect the supernatant for analysis. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Collect the supernatant for analysis, or store at -20°C or -80°C, but avoid repeated freezing and thawing. 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 an aquaporin 4 (AQP-4) 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 aquaporin 4 (AQP-4) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Aquaporin 4 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Aquaporin-4, also known as AQP4, is a water channel protein encoded by the AQP4 gene. AQP4 belongs to the aquaporin family and is an integral membrane protein that conducts water across cell membranes. Its structure consists of six transmembrane domains and five connecting loops forming a channel. X-ray crystallography has revealed that "each AQP4 monomer consists of six helical transmembrane domains and two short helical segments surrounding a narrow water pore." At its narrowest point, the pore measures 2.8 angstroms, just wide enough for single-file passage of water molecules. Although each monomer can transport water independently, the channel's quaternary structure is that of a tetramer. The process by which AQP4 monomers assemble into tetramers is similar to that of other aquaporin channels. Furthermore, it has two distinct structural isoforms in the central nervous system: M1 and M23. The M23 isoform is found as a larger square array on the endoplasmic membrane of astrocytes, while the M1 isoform is smaller and more unstable. Aquaporin-4 tetramers accumulate on the plasma membrane and transform into orthogonal arrays of particles (OAPs). | |||||||||||||||||||||||||||||||||
| 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 | Tissue homogenates, cell lysates, cell culture supernatants, and other biological fluids |
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4.3 ★★★★★
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Product Reviews
★★★★★ 2
Shallow, biased and significantly overpriced
Format: Paperback
Well, this purchase was a disappointment. 20% of the pages are dedicated to just highlighting the bios and backgrounds of the many different authors that contributed this great wisdom. And let me be clear, the authors are solid. They are professionals with credible backgrounds and experience. But it's the format and constraints of this book that makes it virtually impossible for that to shine through. Because the rest of the book (80%) is dedicated to the so called "97 things every cloud engineer should know". And unfortunately the average length of one of these "things" is about 1.5 pages long, and as such extremely shallow and in about 30% of the cases straight up promotions for specific company services. You will find Google cloud advocates telling you to use managed services, of Google of course. AWS engineers telling you to avoid them and use IaaS. LaunchDarkly employees telling you to use feature flags. The list goes on. The TL;DR: here is that if you have built anything on the cloud in the last 2 years, this book is going to be a waste of your time and money. You are better of googling: "cloud best practices" and dedicating 2h to reading the first 10 non-ad related search results.
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Reviewed in the United States on March 23, 2022
★★★★★ 5
Improve Your Relationships
Format: Kindle
In Relationship Goals, Michael Todd has written a book that discussed marriage, dating, and even singleness to help us to grow in our relationships. He wants to help readers to win in relationships and he encouraged us to read the Bible and set goals to improve our relationships. Godly relationships consist of sacrificing for others, displaying kindness, integrity, forgiving others, and loving others. Scriptures declare that we must first love God with all our heart, soul, and mind see Matthew 22:37-38. We are also called to love our neighbors as ourselves (verse 39). He explained how we are supposed to have good relationships with others even if you are introverted. One of the keys, he uses is asking himself does this relationship help me. When we meet the right person, they will help us toward our purpose in life and they will believe in us and love us and they will fit. But if they are moving you away from God, run. He shared how he met his wife when he was 15 years old at a mutual friend’s birthday party. He did everything to make sure she noticed him. This led to a spark and they dated for 8 years except when they experienced an 8-month breakup and he explained what happened. They eventually reconnected and got married in 2010.
I liked how he talked about lot about singleness and how in this time of our life could be the most important time. The reason why is because it’s a time we can focus on what God wants to reveal to us about ourselves and who we are. We are become self-aware and find purpose. This can help us to become whole before we commit to someone else. This is a critical time to heal from our past pains and deal with our fears. You can use this time to get closer to God and getting to know Him.
I would recommend this life changing book to anyone who is ready to improve their relationships it doesn’t have to be just romantic relationships. The same principles can apply to friendships. This book is a very well written book about dating and marriage and how we can change the scope of our relationship. I also liked how he explored the important keys to having a good marriage and what men need and what women needs and the differences. He tries to assist readers in understand each other in marriage and I believe if readers really tried to work on these lessons, they would be less divorce and our marriages would be a stronger example to our children. This book is an awesome book for couples to read and reflect on.
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Reviewed in the United States on January 5, 2025
★★★★★ 5
Written from a Biblical position.
Great read!!!❤️
Excellent for couples who want to create a strong bond and grow.
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Reviewed in the United States on April 1, 2026
★★★★★ 5
Best Relationship Book!!
Format: Hardcover
First, I wish I had this book 25 years ago. Sure, I had the Bible and knew all the Christian 'rules' but Rev. Todd lays everything out so well, even the details that have gotten lost or glazed over through the decades of Christian teachings. The way he presents each phase is superb! I love, love, love his authentic writings on God's love for each of us. You can tell Rev. Todd is a man of deep faith but most of all, he is just a person that, like all of us, mess up.
His writings are real, authentic, relatable, inspiring and exactly what this world needs.
I do hope Rev. Todd continues to write on other topics.
I have been truly blessed by this book!
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Reviewed in the United States on April 4, 2026
★★★★★ 5
Excellent read
Format: Paperback
Great book
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Reviewed in the United States on April 29, 2026