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Description
Mouse prorenin 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 the specimen using EDTA or heparin as an anticoagulant. Centrifuge the specimen at 1000 × g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be assayed or stored at -20°C or -80°C, but avoid repeated freezing and thawing. 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 the tissue and mince it. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1 g of tissue sample to 9 mL 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 the supernatant can be assayed. Cell Lysis Buffer: Gently wash adherent cells with ice-cold PBS, then trypsinize and collect cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash collected cells three times with ice-cold 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 freeze-thaw cycles or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and remove the supernatant for analysis. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, avoiding repeated freeze-thaw cycles. 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 10 ng/mL). Then dilute to the following concentrations: 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, 0.15625 ng/mL, and 0 ng/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 10ng/mL standard working solution into the first EP tube and mix thoroughly to make a 5ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves as a blank well; there is no need to pipette liquid from the penultimate tube. See the figure below for details. 3. Preparation of biotinylated detection antibody working solution: Centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute 15 minutes before use. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration with universal diluent (e.g., 10uL concentrate + 990uL universal diluent). Prepare and use immediately. 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 enzyme conjugate are added sequentially to microwells pre-coated with a prorenin capture antibody. 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 prorenin content in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse prorenin ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Prorenin, a 350-amino acid transmembrane protein, is a precursor to renin. It circulates in blood at concentrations approximately 5 to 10 times higher than renin. It is a hormone that activates the renin-angiotensin system, contributing to blood pressure increases. Prorenin is produced in a wide range of tissues, including the kidneys, adrenal glands, ovaries, testes, placenta, and retina. | |||||||||||||||||||||||||||||||||
| 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 | 0.156-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.9 ★★★★★
Based on 11 reviews
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Product Reviews
★★★★★ 5
I absolutely loved this book.
Format: Paperback
“This book is about creativity, based on histories of contemporary people who know about it firsthand. It starts with a description of what creativity is, it reviews the way creative people work and live, and it ends with ideas about how to make your life more like that of the creative exemplars I have studied. There are no simple solutions in these pages and a few unfamiliar ideas. The real story of creativity is more difficult and strange than many overly optimistic accounts have claimed. For one thing, as I will try to show, an idea or product that deserves the label ‘creative’ arises from the synergy of many sources and not only from the mind of a single person. It is easier to enhance creativity by changing conditions in the environment than by trying to make people think more creatively. And a genuinely creative accomplishment is almost never the result of a sudden insight, a lightbulb flashing on in the dark, but comes after years of hard work.”
~ Mihaly Csikszentmihalyi from Creativity
Creativity with a capital C—the type of Creativity that changes the world.
How can we go about cultivating it in our lives? That’s what this book is all about.
Our guide is Mihaly Csikszentmihalyi—one of the founders of the positive psychology movement. Csikszentmihalyi (pronounced “cheeks sent me high”) is a professor at Claremont Graduate University and the former Chair of the Department of Psychology at the University of Chicago who wrote the classic book Flow: The Psychology of Optimal Experience .
This book features the wisdom gained from over three decades of research along with over one hundred interviews with extraordinary people—from scientists and business leaders to artists and poets.
Warning: Although deeply insightful, it’s not an easy read. If you’re looking to understand creativity at a deeper level via one of the world’s leading (and legendary) psychologists then I think you’ll enjoy it as much as I did.
Here are some of my favorite Big Ideas:
1. What is Creativity - Domain + field + person.
2. Attention - + Being kind weird
3. Complexity - + Resolving dischotomies.
4. Rhythms - Your ideal days?
5. Creativity + Flow - The 9 elements
Well, there we go. That’s a super quick look at this great book. Time to get our flow on as we tap into our Creativity and optimize + actualize!
More goodness— including PhilosophersNotes on 300+ books in our *OPTIMIZE* membership program. Find out more at brianjohnson . me.
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Reviewed in the United States on May 10, 2016
★★★★★ 5
5 Stars – A Must-Read for Anyone Interested in Creativity!
Format: Paperback
Creativity: The Psychology of Discovery and Invention* by Mihaly Csikszentmihalyi is a brilliant exploration of the inner workings of creativity and the human mind. Whether you’re an artist, entrepreneur, or simply someone interested in understanding how creative breakthroughs happen, this book provides a fascinating blend of psychology, real-world examples, and deep insights.
What sets this book apart is its comprehensive approach to creativity, going beyond the idea of it being a mysterious or innate trait. Csikszentmihalyi breaks down the creative process into digestible parts, showing how environment, discipline, curiosity, and flow all play a crucial role in fostering creative discovery. The interviews with a wide variety of highly creative people—including scientists, artists, and business leaders—add incredible depth and context to his theories.
One of my favorite takeaways is how Csikszentmihalyi emphasizes the balance between freedom and structure in creative work, and how finding the "flow" state can lead to moments of true innovation. The book not only helped me better understand my own creative process but also offered practical advice for cultivating creativity in everyday life.
This is an insightful, inspiring read for anyone who wants to dive deeper into the psychology behind creative genius. Highly recommend!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on September 10, 2024
★★★★★ 4
Personal Creativity does not equal Domain Transformation
This was a good if not a great book. Its greatest strength lies in the thesis introduced early on and supported throughout that the kind of creativity that leaves a trace in the cultural matrix rests not in the personal creativity of the individual, but in what Csikszentmihalyi tags the "systems approach " to creativity. To have any effect, a creative idea must be couched in terms that are understandable to others, pass muster with the experts in the field (i.e. the gatekeepers to the domain), and be included within the cultural domain (the set of symbolic rules or procedures) to which it belongs. In this systems view, the definition of a creative person is someone whose thoughts or actions change a domain or establish a new domain (pp. 27-28). This is no easy task, especially since he or she needs to learn the existing domain or domains first, and almost always necessitates being in the right place at the right time (e.g. studying quantum physics at the beginning of the 20th century or women seeking academic opportunities when WWII broke out).
Having established this in the first 30 pages, if you didn't read the remaining 350 you wouldn't miss much. But I still enjoyed reading the stories and thoughts of selected individuals whom the author deemed as "creative" according to the definition above (However, I disagreed with the selection of a few of these and would have chosen at least one more person of faith in addition to the Quaker who was briefly highlighted. Also on the issue of faith, I found the author's grouping on page 371 of studying the bible with addictive behaviors such as cruising the internet and betting on horse races to be rather laughable!).
Some additional personal nuggets I gleaned from this book include the following:
1. Those who persevere and succeed must be creative not only in their manipulation of symbols but maybe even more in shaping a career and a future for themselves that will enable them to survive while continuing to explore the strange universe in which they live (p. 199).
2. When seeking to allow your mind to make new connections in a beautiful setting, just sitting and watching is fine, but taking a leisurely walk seems to be even better. The shaping of one's personal space is also important. The Greek philosophers settled on the peripatetic method, preferring to discuss ideas walking up and down in the courtyards of the academy. When we participate in this kind of "semiautomatic activity" that uses a certain amount of attention, we allow the rest of it to be free to make connections among ideas, often from different domains, well below the threshold of conscious intentionality. "Devoting full attention to a problem is not the best recipe for having creative thoughts. "(p. 138)
3. Both creativity and innovation on the one hand and conservation and traditionalism on the other are both equally important. "Neither uncritical acceptance nor wholesale dismissal of human creativity will lead us far. " (p. 322)
The final section deals with how to enhance personal creativity. Some of these ideas were helpful (e.g. to seek to be surprised and to seek to surprise another person at least once every day, to seek to look at problems from multiple perspectives instead of assuming you see the issue clearly from one perspective, etc.) but others just seem to be taking up space on the page. I'm afraid the phraseology of how to use psychic energy more effectively on page 356 and a few other places lost my interest almost completely.
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Reviewed in the United States on November 15, 2010
★★★★★ 5
interesting analysis of what 'being creative' really means
This easy-to-read absorbing book is based on lengthy interviews with 91 creative individuals ranging from Nobel prize winners to artists to CEOs. Csikszentmihalyi starts by debunking the myth of 'the lone genius having a brilliant idea as if by magic' and defines three necessary ingredients for creativity ('with a capital "C"') - domain, field, and individual. Creativity must take place within a recognised domain (such as physics, painting and so forth); be recognised by experts in that domain (the field, although this may not happen in the individual's lifetime, eg, Van Gogh); and of course come from an individual, although he also adds the painstaking work that precedes and insight, the reality that all creativity builds on what has gone before, and the social elements of the creative process.
The book also offers supporting evidence from the lives of the 91 interviewed, which also provides interesting insights into their lives. In many ways, this book is a biography of the creative individual.
Also contains a chapter with quite practical guidance on how to live more creatively. Prescient advice for a book published in 1996 given the increasing profile creativity is getting in business and public life.
HIghly recommended, one of the most interesting learning experiences i have had in a long while!
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Reviewed in the United States on February 5, 2013
★★★★★ 5
Very enlightening for those who truly appreciate creativity more than cleverness!
Format: Paperback
I love how the author almost redefines creativity .and sheds new light (for me, at least!)on what what real-for-true creativity is and how it benefits individuals and society. It's far more than simply brightening up a room with new wallpaper and curtains -- it describes how genuine creativity requires a thorough working knowledge of the fundamentals of any given field before one can truly create something new or better, and it reveals how those of us who aren't capable of creating something ourselves can yet be part of the process by demonstrating appreciation and support for those who create, whether as sponsors, patrons, or even just ardent fans!
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Reviewed in the United States on July 4, 2023