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Description
Mouse MAS1 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 freeze-thawed. Finally, centrifuge the homogenate at 5000×g for 5-10 minutes, and collect the supernatant for analysis. Cell Lysis Buffer: Adherent cells should be gently washed with pre-chilled PBS, then trypsinized and harvested by centrifugation at 1000×g for 5 minutes. Suspension cells can be harvested directly by centrifugation. Collected cells should be washed three times with pre-chilled PBS and resuspended 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 collect the supernatant for analysis. Other biological fluids: Centrifuge at 1000×g for 20 minutes, and collect the supernatant for analysis. Preparation for the Assay: 1. Remove the reagent kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Prepare a gradient standard working solution: Add 1 mL of universal diluent to the lyophilized standard. Let stand for 15 minutes to completely dissolve, then gently mix (concentration 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 500 μL of universal diluent to each tube. Pipette 500 μL of the 10 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 5 ng/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 MAS1 oncogene (MAS1) 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 MAS1 oncogene (MAS1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse MAS1 Oncogene ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| 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, cell lysates, and other biological fluids |
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4.5 ★★★★★
Based on 1011 reviews
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Product Reviews
★★★★★ 5
What they Didn't Teach You in Sunday School
Format: Paperback
Often when reading through the Bible, whether one does so for religious reasons or not, we miss details in the background. Or we do not take into consideration cultural context while interpreting passages. Sometimes, due to difficulties in translating ancient languages into modern ones, some things are literally lost in translation. In this volume, where Godawa collects material he has previously published into one volume, he examines many of these issues. In particular, he focuses on topics that have long mystified people and been the subject of much conjecture and fictionalization: Nephilim, Watchers and giants. He also takes a close look at verses that may have had supernatural elements inadvertently scrubbed: The strange ariel creatures in 2 Samuel 23:20 translated instead into men, or the demons and goblins of Isaiah 13:21-22 written off as wild animals, etc. Clues to a different ancient world than usually supposed?
The only misstep is his adherence to the John Walton ANE interpretation. This is based on superficial interpretation, and worse history (e.g. the whole ANE "dome" interpretation of creation is largely mythical). Walton's books haves done a huge disservice in undermining biblical inerrancy. See Hugh Ross' Rescuing Inerrancy for more on Walton and others who aren't too good at biblical study.
There's a lot of food for thought in these pages. For more, see
and
.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on September 1, 2018
★★★★★ 5
Great supplement to Bible study
Format: Paperback
Stimulating reading for science-minded and biblically minded readers. The author who wrote this book has a lot to say about some of the strange images we read about in the Bible. Giants, flying reptiles, satyrs, fauns, denizens, angels, demons, etc. Once I read this, I began realizing that much of the strange things we see in the world outside the Bible are about as realistic as some of these images mentioned in the Bible. The battle between our minds and our souls carries on.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on July 26, 2024
★★★★★ 3
Informational but not what I expected.
Format: Paperback
The facts as they relate to the bible and some other scripts are good, but it would have been better if they were backed up with scientific facts and Egyptian findings. This would have given a broader and more open-minded big picture. I was disappointed overall.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 25, 2021
★★★★★ 5
My brain needed this book.
Format: Paperback
What a refreshing cup of water to a thirsty mind! I'm so glad I stumbled across this book and this author, Brian Godawa. I was beginning to think I was the only person on earth who had questions about traditional Christianity versus ancient cultures as well as other mysteries on our earth, and this book has made some links and clarifications on some fascinating subjects that aren't dealt with much in the modern church. It didn't matter that this book is a compilation of appendices based on his research for one of his fiction series. It read, easily, as a non-fiction book and I appreciated how the author used facts based off of biblical and other ancient documents in a rather logical way, with limited conjecture and emotion. I tried absorbing the information in this book as quickly as possible, and already have admired and desired the author's other utterly fascinating book offers. Thank you, Mr. Godawa, for your seeming unrelentless pursuit of truth! I stand with you.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 25, 2021
★★★★★ 5
I own the Canadian copy:
Format: Kindle
This is a captivating and thought-provoking book that dares to explore the uncharted spiritual terrain of the afterlife. Authored with a profound sense of reverence, this book serves as a unique bridge between the iconic voice of John Lennon and the modern reader, yearning for deeper insights into love, life, and the essence of humanity. The narrative is brilliantly crafted, seamlessly blending the ethereal with the tangible. The idea of John Lennon sharing his reflections from Heaven is not just compelling but also deeply moving. The authors have managed to channel Lennon’s timeless philosophy of love, peace, and unity in a way that feels authentic and true to his spirit.
What makes this book truly stand out is its ability to inspire readers to reevaluate their own lives. Lennon’s "heavenly" wisdom challenges us to let go of fear, embrace forgiveness, and lead lives that resonate with harmony and love. It’s a powerful reminder of Lennon’s enduring message and the impact it continues to have on our world. For fans of John Lennon and the Beatles, as well as those curious about the mysteries of life beyond, this book is a treasure trove. It’s not just a read; it’s an experience—a spiritual odyssey that lingers long after the final page is turned.
Whether you're a die-hard Lennon enthusiast or someone simply seeking inspiration, All You Need is Love offers a glimpse into the eternal legacy of one of the greatest icons of our time.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on December 6, 2024
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