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Description
Mouse 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 | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse 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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4.7 ★★★★★
Based on 15 reviews
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Product Reviews
★★★★★ 4
read-this-book-now
Format: Paperback
I liked the pace, the story and the characters. Sadly I found it at the end a bit confusing. I think the book needed more edition work. Otherway, it is a recommendable book if you want horror with a bit of science fiction. Be advised you'll need to use your imagination to understand certain pasages.
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Reviewed in the United States on February 13, 2026
★★★★★ 2
Not even a good read. Pass it.
Format: Paperback
Unfortunately, this book was basically a whole lot of nothing. It was not what I was hoping for, which was on the edge of your seat scary. It was not even alittle scary. Left me with unanswered questions and confused. Sorry..I did not like this book at all.
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Reviewed in the United States on March 6, 2026
★★★★★ 5
Easy to read and fall in love with
Format: Hardcover
one of those books that feels less like a story and more like an experience. Ray Bradbury captures the magic of summer, childhood, and all the little things in life we take for granted. I loved the way it blended nostalgia with those bittersweet moments of growing up. It’s slow at times, but that’s the beauty of it — it makes you stop and notice the small details, just like the characters do.
For me, it felt like stepping back into a simpler time, but with all the emotions and lessons that still matter today. It’s warm, reflective, and beautiful. A book you don’t just read — you feel.
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Reviewed in the United States on September 20, 2025
★★★★★ 5
Vintage Bradbury
Format: Hardcover
Ray Bradbury
August 22nd 1922 - June 5th, 2012
When Ray Bradbury died reactions came from everywhere including from President Obama. Surprising to me, few mentioned the one of his works that meant so much to me and affected my life so deeply. While he was most known to the general public for his science fiction, I found his mostly autobiographical novel Dandelion Wine to be the most impactful. At the same time it best illustrated Bradbury’s incredible command of the language, his ability to stir the imagination, and the way in which he could open windows on life.
I couldn’t count the number of times I would reread a single sentence and become overwhelmed with admiration and envy at how he used words to create images in the mind’s eye. All this was particularly on display in Dandelion Wine and its sequel, Farewell Summer. For Bradbury, it couldn’t be just water. “Nothing else would do but the pure waters which had been summoned from the lakes far away and the sweet fields of grassy dew on early morning, lifted to the open sky, carried in laundered clusters nine hundred miles, brushed with wind, electrified with high voltage, and condensed upon cool air. This water, falling, raining, gathered yet more of the heavens in its crystals. Taking something of the east wind and the west wind and the north wind and the south, the water made rain and the rain, within this hour of rituals, would be well on its way to wine.”
Essentially, Dandelion Wine is the story of a summer in the life of a twelve year old boy as he comes to understand what it means to be alive. But it is also a time capsule for the year 1928 of life in a small town when everyone’s world was much smaller and more compact. There is horror, love, comedy, wonder, nostalgia, and human relations. Bradbury could find unique ways to describe them all.
I first read Dandelion Wine in 1957 when I wasn’t much older than Douglas Spaulding, the central character. It helped me put life in perspective as I was leaving high school. I read it the second time in the early ‘80s when I introduced my daughter to it. Kelly and I sat on our front porch swing one warm summer evening and I read aloud to her the story of Bill Forrester and Helen Loomis. It was all I could do to finish it and when I did we both had tears streaming down our cheeks. Such was the power of imagination and Bradbury’s ability to stroke it to life using just words.
I read it the third time in preparation for reading the sequel, Farewell Summer, written 55 years after Dandelion Wine. Like a fine wine, it had only gotten better with age. Appropriately, Farewell Summer was given to me by Kelly and I read it on summer’s eve 2012. It was the perfect beginning for yet another summer.
In both books the ravine in Green Town, Illinois, based on Waukegan, Illinois where Bradbury grew up was a central feature. I couldn’t resist going to Googlearth to see if the ravine was real. It was. And, it is still there even after Waukegan had changed from a small town to a satellite of Chicago. I was pleased to simply find I could locate it. But when I zoomed in and highlighted the little tree symbol I found the ravine is now Ray Bradbury Park. Perfect!
Dan Winters
June 29, 2012
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Reviewed in the United States on October 24, 2013
★★★★★ 4
One boy’s early awareness of magic and mortality
Format: Kindle
As part of my growing adolescent fascination with the work of Ray Bradbury, of course I read ‘Dandelion Wine’. However, it was one I have not revisited in almost 50 years so my recollection of it is less detailed than many of his other classic books. It’s a collection of interconnected short stories, some previously published, again set in Green Town, Illinois, the fictional counterpart for Waukegan, Illinois where Bradbury spent his first years up until the beginning of his adolescence.
Many of his stories, whether they’re set in Green Town or some other anonymous Midwest town in the 20’s and 30’s resonated with me from the beginning. My father was born just a few months after Bradbury and grew up during that same time in another small town in Missouri, which I recall visiting a few times in my childhood and seeing a neighborhood not much different from Bradbury’s, and a house almost literally unchanged from the time when my father was a boy.
That nostalgia, that yearning for the freshness and intensity of a child’s perception, when a boy will find magic in a birdbath and an earth-scented basement, definitely spoke to my soul and still does, 50 years later.
The main character is a Ray surrogate, a twelve-year old boy named Douglas Spaulding (Bradbury’s middle name is ‘Douglas’) who has a ten-year old brother named Tom. They live with their parents, grandparents, and great-grandmother in an old house that is sturdy and roomy enough to accommodate a few boarders. One of the ‘beginning of summer’ rituals is the bottling of dandelion wine that will last the entire summer and beyond, at which point it will be a way of preserving what was memorable about the summer that just passed.
‘Hold summer in your hand, pour summer in a glass, a tiny glass of course, the smallest tingling sip for children; change the season in your veins by raising glass to lip and tilting summer in.’
During this particular summer, Doug fully realizes, for the first time, that he is alive and, conversely, that he will die. He holds mortality at bay as much as he can, with special sneakers in which he can run from one end of the town to the other and working out a clever bartering trade with the shoe salesman as a way to “buy” the sneakers. Doug could be a future salesman himself, persuading the salesman to try on a pair himself so he will know what he’s selling and how it actually feels to wear a pair. The future writer Doug also wants to document every significant event that happens to him this summer of 1928.
His younger brother Tom, on the other hand, is more logical and reasonable. While Doug chronicles the events of the summer, Tom records data such as the first rainfall and other meteorological data. Tom also seems to me to be the wiser of the two, reasoning with and calming down the melodramatic Doug on more than one occasion.
Everything in the town acquires new meaning to the otherwise carefree and playful Doug. There are discernible boundaries between civilization and wilderness in this little hamlet, the most notable example being the ravine:
‘The ravine was indeed the place where you came to look at the two things of life, the ways of man and the ways of the natural world. The town was, after all, only a large ship filled with constantly moving survivors, bailing out the grass, chipping away the rust.’
The death of his great grandma also occurs this summer. After a lifetime of activity and housekeeping and family keeping, she decides that she has lived long enough. She has no discernible ailment, just a “mild but ever-deepening tiredness”. She has to assure Doug and Tom that the time for doing all this activity has come to an end and that they must learn to accept it.
Just as disturbing for Doug is when his best friend John Huff tells him that his father is being transferred to Milwaukee .His family is leaving on the train that evening. John is a budding young superman. He is a master pathfinder, swimmer, climber and jumper. He is also not a bully. He is kind as well as smart. As far as Doug is concerned, he is a god. For their last play activity, they play a game of hide-and-seek. Doug volunteers to be ‘it’, hoping by controlling the pace of the game to prolong John’s departure. John wraps that one up and agrees to play one more game, with him as ‘it’. With Doug and the other boys frozen into ‘statues’, John punches him on the arm gently, saying “So long” and then runs.
There is even a serial killer in Green Town, referred to as The Lonely One. Young spinster Lavinia Nebbs and some of her friends are worried about the disappearance of another of their friends. Rumors of the Lonely One being on the loose abound with the deaths of two young women occurring within the past two months. With the disappearance of their friend they have ample reason to be concerned. Then they find her, lying dead on the ground. They find the police and, after he finishes questioning them, they are free to leave. Lavinia, putting on a brave front, suggests they go to a Charlie Chaplin movie to stave off their fear. This works pretty well until the film ends, the last feature of the night, and they all have to walk home in the dark. Lavinia, still trying to hide her fear behind a brave front, agrees to walk her friends home first, meaning that she’ll have to walk the rest of the way to her house by herself. Bradbury’s mastery of suspense is particularly evident in this chilling and terrifying episode. I won’t reveal the outcome.
There is one episode in which Doug and Tom, primarily Doug, come to believe that a wax, fortune-telling “Tarot Witch” automaton is actually a mummified queen from ancient Egypt. In reality it is a slot machine in which you put in a penny and out comes a card with your fortune written on it. The alcoholic owner is disgusted with it and his failing slot and pinball machine business and ready to throw it in the trash heap. Doug and Tom attempt to rescue it. This sequence is long and tedious and has the effect of Tom and Huck rescuing Jim near the end of ‘Huckleberry Finn’. In both cases it’s an unwelcome diversion that detracts from the power of the novel.
Overall, ‘Dandelion Wine’ works. It is not as disjointed as it seemed to me 50 years ago when I could detect the short story origins of much of it. Depicting the course of a summer is by its nature episodic. There are moments where it seems that everybody talks like Bradbury writes, even the semi-literate characters, and with a zeal and enthusiasm that gradually took over most of his later fiction. At its core, however, it captures, through a poetic filter, the magic and intensity of a child’s perception and his awareness that all this beauty surrounding us is fleeting so we may as well appreciate it as much as we can while we can.
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Reviewed in the United States on May 10, 2022