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Description
Human NPHN 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: Urine 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
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: Urine 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 utilizes a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA). Sample, standard, biotin-labeled detection antibody, and HRP enzyme conjugate are sequentially added to microwells pre-coated with a nephrin (NPHN) capture antibody. 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 nephrin (NPHN) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Nephrin ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Nephrotic hormone (NPHN) is a protein essential for the proper functioning of the kidney's filtration barrier, which is composed of slit endothelial cells, the glomerular basement membrane, and the epithelial capsule. It is a transmembrane protein and a structural component of the slit diaphragm. It is present in an intricate network at the apical end of the capsule cells, imparting a strong negative charge that repels proteins from passing through Bowman's space. Defects in its gene, NPHS1, are associated with Finnish congenital nephrotic syndrome, resulting in the leakage of large amounts of protein into the urine, a condition known as proteinuria. It is also essential for cardiovascular development. | |||||||||||||||||||||||||||||||||
| 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 | Urine, tissue homogenates, cell lysates, cell culture supernatants, and other biological fluids |
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4.1 ★★★★★
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Product Reviews
★★★★★ 5
-Daughter of Amazon Account-
Size: 3 Fl Oz (Pack of 1)
In the past I have used high end skincare products and wasted money. This is absolutely one of the best PM OIL-FREE facial moisturizers. This has helped my skin improve significantly over the last few months. Its super super light weight and pea size dots cover my entire face. I add a pea size to my cheeks, forehead and chin. It all soaks into my pores evenly without clogging them. Due to me using a serum by CeraVe as well.
(Very important part of my personal skin-care routine)
I have extremely sensitive skin and nearly break out with everything I use.
However this helps being fragrance-free & OIL-FREE!
Over the years I always thought high end brands were better than the lower cost friendly ones. (NOT THE CASE! At least for me.)
I highly recommend this product for someone who is starting out using facial products or someone who has indulged in high end.
This does WORK! And it leaves my skin feeling fresh and lightweight. I started seeing differences in my skin texture (for the good) within about a month after using it daily. I then backed off from it and use it every other day. Since I now have my skin where I need it to be.
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Reviewed in the United States on April 11, 2026
★★★★★ 5
LIGHTWEIGHT
Size: 3 Fl Oz (Pack of 1)
I LOVE THIS.IT'S DEF LIGHTWEIGHT YOU CAN FEEL IT AND SO MOISTURIZING,EASY TO USE AND A LITTLE GOES A LONG WAY.THE CONSISTENCY IS AMAZING AND DOESNT HAVE A FRAGRANCE.I GUESS ITS WORTH THE PRICE BUT NOT SO AFFORDABLE OPTION COZ OF THE QUANTITY.I NEVER REACTED SO GOOD FOR SENSITIVE AND ACNE-PRONE SKIN
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Reviewed in the United States on May 11, 2026
★★★★★ 5
Affordable and Effective
Size: 3 Fl Oz (Pack of 1)
This is a great everyday moisturizer for before bedtime. It is a bit heavier than some moisturizers, but I personally like that at night. My skin seems to drink it up as I sleep and in the morning I wake up without any puffiness to my face. I look refreshed and it never feels greasy or sticky. Best part is the price. It is so affordable.
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Reviewed in the United States on May 23, 2026
★★★★★ 5
Remember: Margin of safety
Format: Paperback
Why the book was so easy and enjoyable to read? It has a lot of good examples, data, and fun facts to get the point across to the readers. The chapter titles are attention grabbers that get our attention so that we can read more. However, the most important thing to learn from this book is the "Margin of Safety."
According to the author, it is one of the most underappreciated forces in finance. It comes in many forms: a frugal budget, flexible thinking, and a loose timeline - anything that lets you live happily with a range of outcomes. Controlling your time is the highest dividend money pays.
The book is pretty much evolved around the concept of "Margin of Safety." It encourages readers to save money and not spend money lavishly. The key is staying wealthy and not just getting wealthy. We can't be complacent and assume that yesterday's success translates into tomorrow's good fortune. Wealth is what you don't see. Spending money to show people how much money you have is the fastest way to have less money.
Good investing is not about getting the highest returns. It's about getting good returns that you can stick with and which can be repeated for the longest period of time. According to the author, the historical odds of making money in US markets are 50/50 over one-day periods, 68% in one-year periods, 88% in 10-year periods, and (so far) 100% in 20-year periods.
Forecasting is hard. This is why investment guru Benjamin Graham strongly advocates for the margin of safety, as the purpose of the margin of safety is to render the forecast unnecessary. The author cited the success rate of venture financing from 20024 to 2014: 65% lost money, 2.5% of investments made 10X to 20X, 1% made more than 20X return, and only 1/2% (~100 companies) earned 50X or more.
According to George Soros, it is not important whether you are right or wrong but how much money you make when you're right and how much you lose when you're wrong. You can be wrong half the time and still make a fortune.
The most interesting part of the book is the last chapter: Postscript. Thanks to the internet, the world is more connected than ever. That means that the talent pool the readers compete with has gone from 100s or 1000s sprang their towns to millions or billions spanning the globe.
The author ended the book with a not-so-pessimistic note. The era of "this isn't working" may stick around. And the era of "We need something radically new, right now, whatever it is" may stick around.
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Reviewed in the United States on February 25, 2024
★★★★★ 5
The Psychology of Money: A Masterclass on Wealth, Human Nature, and True Happiness
Morgan Housel’s The Psychology of Money is not your typical finance book. It's an insightful and profound exploration of how human behavior, rather than cold hard numbers, often determines financial success—or failure. If you’re looking for a book that teaches you how to manage wealth, understand greed, and find happiness, this is a timeless treasure trove of wisdom that transcends spreadsheets and stock markets.
Lessons in Human Behavior, Not Just Finance
Housel's genius lies in his ability to connect finance to human psychology, showing how our emotions, biases, and decision-making habits influence our financial outcomes. Unlike most personal finance books that focus on technical advice, this one delves deep into the mindset required to build and maintain wealth. Through engaging storytelling and real-life anecdotes, Housel illustrates that how we think about money is often more important than what we actually know about it.
The Power of Compounding Behavior
One of the book’s core messages is the immense power of compounding—not just in terms of investments but in life itself. Housel masterfully explains how small, consistent decisions can lead to huge gains over time, whether in wealth-building, relationships, or personal growth. He reminds us that patience and discipline are the cornerstones of financial success, and that short-term thinking is often the enemy of long-term wealth. His examples of how figures like Warren Buffet amassed fortunes through simple, disciplined investing make this concept strikingly clear.
Greed: The Silent Wealth Killer
Greed is one of the most destructive forces in personal finance, and Housel addresses it head-on. Through stories of financial bubbles, crashes, and personal downfalls, he shows how the relentless pursuit of "more" can derail even the most secure fortunes. His exploration of why it’s so hard for people to "have enough" is a sobering reminder that wealth is as much about mindset as it is about numbers. The book doesn’t just highlight the dangers of greed; it also offers practical ways to avoid falling into its trap by cultivating a sense of financial contentment.
Happiness Beyond the Dollar Signs
While the title suggests that money is the focus, happiness is the true heart of this book. Housel argues that wealth, when viewed properly, is a tool for freedom rather than a scorecard. His chapters on the importance of controlling your time, living below your means, and the intangible rewards of financial security are powerful reminders that happiness isn’t just about how much you earn, but how well you live. He masterfully weaves together the idea that wealth is not the end goal, but a means to achieve a life filled with joy, autonomy, and purpose.
Timeless Lessons for Every Reader
What sets The Psychology of Money apart is its universal appeal. Whether you're a seasoned investor, a financial novice, or someone simply seeking a healthier relationship with money, the book’s lessons are relevant and accessible. Housel’s conversational writing style makes complex concepts feel straightforward, and his ability to blend financial advice with psychology and philosophy makes this book a must-read for anyone wanting a holistic approach to money and life.
Final Verdict: A Wealth of Wisdom
Morgan Housel’s The Psychology of Money is a masterpiece of personal finance and self-awareness. Its lessons on wealth, greed, and happiness go far beyond dollars and cents, challenging readers to rethink their relationship with money and life itself. This book isn't just about getting rich—it’s about getting smart, getting wise, and getting happy. A timeless, essential read for anyone looking to master not just their money, but their mindset.
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Reviewed in the United States on September 5, 2024