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Description
Mouse DEFa6 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: Serum, plasma, 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
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: Serum, plasma, 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 1000 pg/mL). Then dilute to the following concentrations: 1000 pg/mL, 500 pg/mL, 250 pg/mL, 125 pg/mL, 62.5 pg/mL, 31.25 pg/mL, 15.625 pg/mL, and 0 pg/mL. Serial dilution method: Take seven EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 1000 pg/mL standard working solution into the first EP tube and mix thoroughly to make a 500 pg/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 Defensin Alpha 6/Paneth Cell Specific (DEFa6) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase and to yellow by acid. The intensity of the color is positively correlated with the amount of Defensin Alpha 6/Paneth Cell Specific (DEFa6) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Defensin Alpha 6; Paneth Cell Specific ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Defensin α6, Paneth cell (DEFA6) is a protein encoded by the DEFA6 gene. DEFA6 is expressed in Paneth cells of the ileum. α-defensins are a family of microbicidal and cytotoxic peptides that protect the host against bacteria and viruses. However, DEFA6 provides protection against invading enteric bacterial pathogens by self-assembling into cellulose and nanomembranes that surround and entangle bacteria. Several α-defensin genes, including DEFA6, are clustered on chromosome 8. | |||||||||||||||||||||||||||||||||
| 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 | 15.6-1000 pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate and other biological fluids |
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4.9 ★★★★★
Based on 1508 reviews
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Product Reviews
★★★★★ 5
Best Book on the Integration of Faith and Learning
Format: Paperback
A problem area in Christian ministry is the area of Christian higher education. As we continue to progress through the 21st century we continue to see the decline of the Christian higher education movement. What was once a strong area in the Christian ministry, Christian higher education is failing. The Bible College movement has been in decline for sometime. Schools are folding without the students or the funds to stay open. Most people are going to secular colleges and universities over Christian schools. One of the major problems with Christian higher education has been the failure to critically interact with the movement and offer an approach to dealing with this decline. David Dockery has helped fill this void with his recent volume, Renewing Minds. Dockery, President of Union University in Jackson, TN, is extremely qualified to write in this capacity. A clear and thoughtful theologian, he has extensive experience in the areas of leading and administrating a Christian higher education institution. Not only has he lead Union University he also serves as chairman of the board of the Council for Christian Colleges and Universities. With recommendations from J. I. Packer, R. Albert Mohler, Chuck Colson, and a foreword by Robert P. George of Princeton University, this is a volume that should be seriously considered by all who love Christian education.
In Chapter 1, Dockery highlights the problem in America. He writes,
"I believe that the integration of faith and learning is the essence of authentic Christian higher education and should be wholeheartedly implemented across the campus and across the curriculum. This was once the goal of almost every college in America. This is no longer the case.... What happened was a loss of an integrated worldview in the academy. There was a failure to see that every discipline and every specialization could be and should be approached from the vantage point of faith, the foundational building block for a Christian worldview" (pp. 5-6).
Tracing the history of the departure of American schools into secularism and surveying the kinds of Christian higher education institutions in North America leads to a defense of the system derived from Matthew 22:36-40 and the Great Commandment to love the Lord your God with your mind! The rest of the book explains how to go about obeying the Great Commandment in Christian higher education. Chapter 2 builds on this by explaining from the Scriptures the role of the Christian higher education institution and deals especially with the role of the Church, and therefore the Christian higher education institution in society. Chapter 3 explains the process of shaping a Christian worldview and the impact on this on Christian higher education. Chapter 4 is about reclaiming the Christian intellectual tradition. Dockery writes here after tracing the history of the Christian intellectual tradition
"Certainly we all learn apart from the great Christian intellectual tradition, apart from the vantage point of faith. But we cannot connect these things into a unified whole, we cannot fully understand the grand metanarrative; we cannot truly grasp how to explore and engage the issues in history and science, business and health care, apart from this approach to learning. Thus we must seek to sanctify the secular because Jesus Christ has come to earth" (p. 84).
Chapter 5 addresses the issues of integrating faith and learning. Chapter 6 addresses the necessary concept of developing a place of belonging and community where scholars, educators, staff, and students live together, share, serve, and learn. Chapter 7 begins to offer practical ways of establishing this grace-filled academic community. Chapter 8 articulates how to develop a theology of Christian higher education. Developing this theology would have positive implications for the academic community and the individual. Chapter 9 serves as the culmination of the book with thinking globally about the future. With the changes in communication we must embrace the new in order to communicate the orthodoxy of the past into a new global world. This means listening as much as talking especially as global Christianity begins to reflect non-Western images, positions, and principles. Christian higher education does not just simply say the West is best but listens to all Christian voices in order to best communicate the timeless truth in new ways. This is then concluded by an extensive bibliography on the integration of faith and learning.
Dockery's book fills a great need in the area of Christian higher education. He states the issues and the problems, traces the history of Christian higher education, articulates a biblical defense of the integration of faith and learning as well as a comprehensive theological defense. Not only does he articulate this at an academic level but he does not neglect the spiritual aspect of things, emphasizing not just "smart" Christians but "spiritual" Christians. The movement from "theory" to "practice" in Dockery's book is exceptional. I hardly find anything in it that I would disagree with or anything I wish I say that I did not see in the book. It is an even handed treatment that should be read by those who care about Christian higher education and especially those involved in Christian higher education. May we see a renewal of a close integration of faith and learning on our campuses as we emphasize the great truth that all truth is God's truth. May we raise up godly men and women who are passionate about the truth and about serving Christ in the world around them through the Great Commission. And may those of us involved in Christian higher education lead the way through authentic spirituality grounded in the truth. Highly recommended!
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Reviewed in the United States on October 10, 2009
★★★★★ 2
Not much about higher education
Format: Paperback
I gave this book 3 stars not because I think it was bad, but because it didn't really have much to do with higher education. I am a big believer in Christian higher education and the integration of faith and learning, however, if you were to take this book and replace "Christian higher education" with a phrase like "the Christian community" or the "Church family" no one would notice the difference.
I do believe in much of what he said but that's because I follow Christ. I didn't expect him to spend chapters on what Christians believe and how they differ from other religions, I was hoping for an intelligent argument and exploration of Christian higher education and how it differs from other higher education. And the argument, higher education used to be all Christian higher education is not a good argument.
Once again, not a bad book but just not what I expected based on the description and title.
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Reviewed in the United States on December 7, 2011
★★★★★ 5
A Sterling Vision of Christian Education
David Dockery is the president of my alma mater, Union University in Jackson, Tennessee. Therefore, I have always taken great interest in keeping up with what Dockery says and does in the realm of Christian higher education. B&H publishing has done us all a favor by pulling together his ideas into a unified book with the theme - "Serving Church and Society through Christian Higher Education".
Dockery's heart beats with the passion of a pastor, theologian, academic, and administrator. He sees the Christian university as a place in society where both mind and heart can renewed along biblical and gospel lines. It is difficult work in our day, but it is a necessary work.
Dockery writes, "I believe that the integration of faith and learning is the essence of authentic Christian higher education and should be wholeheartedly implemented across the campus and across the curriculum."
And how is this accomplished? Dockery says, "We need more than just new ideas and enhanced programs, we need distinctively Christian thinking, the king of touch-minded thinking that results in culture-engaging living. ...This perspective involves the whole of our human personality. Our minds are to be renewed, our emotions purified, our conscience kept clear, and our will surrendered to God's will. Applying the Great Commandment entails all that we know of ourselves being committed to all that we know of God."
A number of the chapters in this book simply sparkled with insight. Pastors will especially note the overlap of Dockery's vision of Christian community in the university with what we also hope to find within the local church. For example, Dockery writes a chapter on "Establishing a Grace-Filled Academic Community" that could and should be applied to the local church as well, with an emphasis on unity, shared life, worship, and service. Within chapter six is a section titled, "Building Blocks for Building a Community with Renewed Message", a message with such urgency and clarity that I did in fact bring it home to our church for a renewed sense of Christian community.
Such is the case for much of this excellent book. You may not have a vocational calling to higher education. However, as a pastor or Christian parent, it is your responsibility to consider carefully the type of institution you send your students to for university education.
Dockery writes, "I would suggest that the starting point of loving God with our minds, thinking Christianly, points us to a unity of knowledge, a seamless whole, because all true knowledge flows from the one Creator to His one creation." Dockery's vision is compelling and sound, and I heartily recommend this book.
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Reviewed in the United States on November 22, 2007
★★★★★ 5
Good Value & Good Product.
For those of us that don't eat a lot of fruits and veggies normally, this product really helps. It meets my needs for fruits and veggies. It's easy to take, goes down well, and has no after taste. Good value too.
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Reviewed in the United States on April 11, 2026
★★★★★ 5
Good product, reasonable price.
Good product. Easy to swallow. Reasonable price.
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Reviewed in the United States on May 22, 2026