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Description
Human PTTG1IP 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 ultrasonically disrupted or repeatedly frozen and thawed. Finally, the homogenate is centrifuged at 5000×g for 5-10 minutes and the supernatant is collected for analysis. 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 conjugate are sequentially added to microwells pre-coated with a capture antibody against Pituitary tumor-transforming gene 1 protein-interacting protein (PTTG1IP). 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 amount of Pituitary tumor-transforming gene 1 protein-interacting protein (PTTG1IP) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Pituitary tumor-transforming gene 1 protein-interacting protein ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Pituitary tumor transforming 1-interacting protein (PTTG1IP), also known as PTTG1-binding factor (PBF), is a protein encoded by the PTTG1IP gene. It is a poorly characterized protein. Despite its ubiquitous presence in normal tissues, its precise function remains elusive. It has been shown to directly interact with the safety protein and proto-oncogene PTTG1, promoting its nuclear translocation and subsequent transcriptional activation of basic fibroblast growth factor (bFGF) by PTTG1. It may have a direct role in cancer. It has been shown to regulate thyroid cell growth, with overexpression leading to thyroid hyperplasia and the formation of intrathyroidal lesions. Its expression is also independently associated with tumor recurrence. It has also been implicated in breast cancer. Its overexpression and secretion induce cell invasion, a process crucial for the development of metastatic disease. | |||||||||||||||||||||||||||||||||
| 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 | Tissue homogenates and other biological fluids |
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4.4 ★★★★★
Based on 30 reviews
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Product Reviews
★★★★★ 4
The Best of the Modern Guides to the PCT.
Format: Paperback
This book is not an update of the Wilderness Press classics on which the trail was discovered and hiked by a couple generations from the 1970s to around 2010. It does however owe those books a heavy and unacknowledged debt. This is most notable in lettering sections which the author of this guide, Shawnte Salabert, attributes to the PCTA but in fact come from scarified, Schaffer, et all who wrote the original guides to California, Oregon, and Washington. Even the definition of sweat constitutes the Southern California trail, comes from those guides. (It starts at the Mexican Border near Campo California and ends in Tuolumne Meadows, Yosemite.). But alas, those books are long out of print, in part because today’s hikers prefer light weight apps with minimal trail information as opposed to detailed descriptions of the physical geography and varied ecosystems of the trail. I’m not sure the lost knowledge has equaled the reduced weight but I’m a bit old school.
And this book is as much a throwback to that kind of writing as it is a nod to recent demands of the hiking community. It does cover every mile of the route, suggest campsites along the way, and as much as possible provide an option for day and section hikes along the trail for just about anyone. The author provides good car descriptions to major trailheads for each section hike he recommends as well as available entry and exit options along the way. The writing is entertaining and the author has his own sense of humor that will become readily apparent as you read the text. Based on my experiences (and I’ve hiked 95% of the trail he describes at least once; sometimes multiple times) his mileage descriptions are accurate and you will easily be able to recognize the places he describes along the way. The book also features nice color photos and it’s availability in digital form will certainly please the gram counting hiker set. On the other hand, I miss seeing elevations along with miles at each major trail intersection, pass etc. Salabert does give total elevation gain and loss for each section of trail he describes but sometimes it’s nice to know just how much of a climb to expect.
Ultimately, this book fulfills it’s purpose. You should want to hike the PCT after reading a few pages if you hadn’t already when you purchased the book. And if you are like many people with only a weekend or a few days to spare hiking the trail, this book will make planning short sections easier. It offers a lot. But it reminds me of an era when guidebooks offered even more.
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Reviewed in the United States on June 17, 2020
★★★★★ 5
Superb Guide in All Respects
Format: Paperback
An invaluable reference for anybody interested in section hiking the trail. I especially appreciate the coverage of water and campsites. It’s also a fun and beautiful read for any armchair adventurer. Salabert did an amazing job bringing together well-written text, data, maps, and beautiful photos to create a comprehensive guidebook. Deserves to do very well.
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Reviewed in the United States on February 6, 2018
★★★★★ 5
Very good comprehensive collection of Section Hikes of the PCT
Format: Paperback
Great Maps. Accurate descriptions of everything (verified on previous hikes and hikes using the book).Well organized and beautifully laid out.
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Reviewed in the United States on August 28, 2021
★★★★★ 5
Truly embracing life along the Camino de Santiago
Format: Paperback, Format: Paperback
The Way of the Wind: Embracing Life While Walking the Camino do Santiago, by John W. Pearson, 2021, recounts one man’s experiences while walking the Camino de Santiago, a thirty-five day, five hundred mile walking journey from St. Jean Pied de Port in France to Santiago de Compostela in Spain. The journey follows a path tread by literally millions of pilgrims, or “pelegrinos”, as they seek to find something very person in the experience. The book is a travelogue recounting the miles of the daily trek, the people, stories, dining, lodging, and local services encountered along the way. There are a series of black and white photographs accenting the dialogue which give the reader a sense of involvement in the story. A nice touch was a simple map at the beginning to orient the reader along each day’s travel.
I absolutely loved this book. It was enjoyable to read and I found myself slowing down near the end of the book to better savor the details. It warrants a double read to go back and let those early experiences soak in a bit more.
I found the book to be educational, as I learned a lot about the historical significance of the route of the Camino, as well as specific landmarks along the way, such as a 10th century Roman bridge dwarfed by a new modern bridge overhead. I imagined myself walking along that ancient roadway, imagining what a pilgrim in 1300 must have thought.
The dialogue is playful, recount with hours of walking, lively dinners, and crowded “albergues” (think camp dorm rooms with bunk beds).
All was not rosy along the Camino. The narrative relates the aches of sore feet, legs, backs; the challenges of securing lodging and laundry; meanings lost in translation; inclement weather and treacherous roadways. Often mentioned are markers to honor the fallen dead along the Camino.
About 2/3rds through the book, John recants his very personal reason for going on the pilgrimage, which he describes as “The Whale in the Room”, referring to the motivation of Captain Ahab in Moby Dick. His honesty and vulnerability bring the entire journey into focus. He brings the reader to the Cruz de Ferro, where pilgrims have stopped for millennium with their deepest meanings.
I have known John Pearson since 1975, when we met in High School in El Paso, Texas. However, through the years, I moved and we lost close touch. There were many parts of this book of which I did not know the details. John’s story is very powerful, and will impact the reader deeply.
Once you have read the book, you will have a clear idea of the experience, and can judge for yourself if the Camino de Santiago is calling you. For me, I found the book asking me not “WILL you go?”, but “WHEN will you go?”
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Reviewed in the United States on May 9, 2021
★★★★★ 5
A remarkable story, illuminating and real.
Format: Paperback
This book is a journey, the author’s journey and potentially the reader’s as well. A story that at once reveals the author’s walk of hundreds of miles, intertwined through steps of ancient history, beautiful and interesting places, and traversed by countless people who through the ages embarked on similar journeys of their own. There is a revealing honesty that comes through these pages allowing the reader to feel vicariously on a kind of journey of one’s own. With great admiration for this writer and for all those who have traversed so consummate a pilgrimage, I found myself searching my own soul, visualizing the vast and beautiful detailed descriptions, enjoying the frequent humor and occasional hilarity, at times laughing out loud, feeling deeply moved, filled with questions and ideas about the ways we all find ourselves traveling our lives. The people, fellow pilgrims the writer meets along the way, the experiences they share on their subjective journeys, are honored by this author as are the lives of those who came before them over the centuries. It is a book filled with surprises, joy, pain, beauty. An absolutely awesome experience to read.
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Reviewed in the United States on July 8, 2021