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Description
Rat AQP3 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, biotinylated detection antibody, and HRP conjugate are sequentially added to microwells pre-coated with a capture antibody against Aquaporin 3, Gill Blood Group (AQP3). After incubation and washing, the sample is developed using the substrate TMB. TMB converts to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of Aquaporin 3, Gill Blood Group (AQP3) 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 3, Gill Blood Group ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Aquaporin-3, the protein product of the AQP3 gene, is present on the basal cell membrane of the primary collecting duct cells, providing a pathway for water to leave these cells. Aquaporin-3 is also permeable to glycerol, ammonia, urea, and hydrogen peroxide. It is expressed in various tissues, including the skin, respiratory tract, and kidneys, as well as in various types of cancer. In the kidney, aquaporin-3, unlike aquaporin-2, does not respond to vasopressin (also known as antidiuretic hormone). | |||||||||||||||||||||||||||||||||
| 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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4.8 ★★★★★
Based on 21 reviews
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Product Reviews
★★★★★ 4
70% practical advice, 15% feel-good advice, 1% useless advice, 14% advice that saved my academic life
Format: Paperback
To be honest, if I had just graduated high school and someone gave me this book as a gift, I would roll my eyes and never open it. I picked it up a week ago, however, now that I am almost done with my second year of university, and I really do wish that I had had it (and cared to read it) two years ago.
The book is seventy-five pieces of advice, each with about two pages of explanation. The advice is pretty simple, as you can see from looking at the table of contents ("Dress Nicely for Class," "Never Nap," "Eat Healthy," "Always Go to Class"). But the reason this book is effective is that it serves as a quick-reference manifesto for some of the more important (to me) advice.
For example, I'm taking a grad-level fiction writing class. No due dates (except the final deadline at the end of the semester) and no class. You just write at your own pace and turn in a portfolio. This is incredibly difficult for me to do, and I'm unbelievably far behind in the work for the class. I was really quite worried about how I would ever pull it off. The whole semester, my fiction work has been priority #75, and I usually crash between priorities #14 and #20. But with some of the advice in this book ("Keep a Work Progress Journal," "Set Arbitrary Deadlines," "Avoid Daily To-Do Lists," "Don't Take Breaks Between Classes"), I actually feel pretty confident about being able to finish on time. By reading this book (and [...] and Newport's more recent book,
) I've adopted some strategies and habits that have actually yielded results.
Before Cal Newport, I was up late every night, angrily doing my homework until I couldn't drink any more tea, without any free time. Now, I'm getting my homework done before sundown (for the most part), feeling enormously more relaxed, and regaining a good amount of the excitement that I had about college before I got here.
If you're on your way to college, and you're the sort of person who can stomach (and listen to) advice, do yourself a favor and read this book. Newport admits in the introduction that not every piece of advice will be for you (for me: "Exercise Five Days a Week" and "Use a Filing Cabinet"). If you want to be more than an average student without being a "grind," this book has a good deal of solid advice.
If you're already in college, and you're looking for more in-depth and practical advice, I recommend also reading
. It has more thorough advice for confronting the terrors that you have come to know in college.
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Reviewed in the United States on April 8, 2010
★★★★★ 5
Will Help You Reach the Goal You Thought You Could Never Achieve
Format: Paperback
Quick review: if you're about to go into college, BUY THIS. If you're already in college, STILL BUY THIS.
This great book has conventional and unconventional advice. I'm already a fan of Cal Newport's advice and blog, but this book condenses his ideas that are most relevant to me (since I will be going to college in the fall of '15).
It isn't a typical 'how to do well in college' book whatsoever. This truly gives you the truth of what needs to happen to become a part of the exceptional. And he isn't writing it with the assumed purpose of impressing everybody or becoming a mega-successful overlord; he writes with the tone that leaves up the ultimate goals you have up to you. Also, the writing style is not annoying like a lot of books are.
Personally, I just want to optimize my college experience, leave my comfort zone, and get into grad school. This book will help me.
If you want to start the track of becoming a super-rich, art-gallery-going, Bentley-driving, awe-inspiring successful person, this book will still be very worth your time.
It's absolutely worth the money if that's what is keeping you on the edge of buying this.
It has countless (actually 75) useful ideas throughout, too, if you worry it has only one or two pieces of useful advice and that's it.
Bonus: if you like what you read, he has a blog with even more stuff to learn.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on August 5, 2014
★★★★★ 5
Fantastic Advice
Format: Paperback
I'm a senior in high school preparing for my freshman year.
This book is amazing. The advice is down to earth and believable without being preachy or perfect. Newport is perfectly willing to admit that things aren't always going to go exactly as planned, but still tells his readers how to accomplish great things. I absolutely loved the advice. Some of the advice in this book is not for everyone - APPLY TO TEN SCHOLARSHIPS A YEAR can be overwhelming and unnecessary, depending on the student. That said, it's easy to see how even applying a handful of these tips would change my college career for the better. As a student who will have to balance work, class, and a social life, I think this will be rewarding. Here are a handful of my favorite tips:
CREATE A SUNDAY RITUAL
STUDY IN FIFTY MINUTE CHUNKS
USE THREE DAYS TO WRITE A PAPER
EAT HEALTHY
VOLUNTEER QUIETLY
And the list goes on. I will definitely pack this with me in the fall.
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Reviewed in the United States on April 11, 2010
★★★★★ 5
Get it and Use it!!
Format: Paperback
Did not get to attend an Ivy or a Top 20 / 30 / 50 most selective University / College?? - But still want to make sure you don't lose out in comparison with those students who did?? Then go get yourself a physical copy of this book - read it, use it - choose some strategies and implement it. If you did - you will ensure that you maximized your college experience and did not lose out because you didn't land up going to those selective colleges. And even if you are in those selective colleges - you can use this book to maximize the value of your opportunities during your stay at those Universities. Good Luck in your quest!!!!
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Reviewed in the United States on August 5, 2025
★★★★★ 5
Wise Advice for the College-Bound Student
Format: Kindle
I read this book after reading Newport's other study guide How to Become a Straight A Student and was looking forward to more of his easy to understand, quickly read nuggets of wisdom. Although I found the advice in his previous book to be more helpful, I did fine some words of advice that I will surely put to use in my upcoming semester.
Among the gems I found, Newport reminds students to "Study in Fifty-Minute Chunks" and "Learn to Listen." In these short chapters there are ideas to help the student better utilize their time by minimizing distractions and knowing exactly what to listen for in a lecture. By far my favorite and most memorable line from this book is "A good analogy is that writing is to a college student what shooting hoops is to a basketball player." Wise advice indeed. I would recommend this book to high school an college students alike. The chapters are short and jam packed with good advice.
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Reviewed in the United States on June 25, 2012