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Description
Mouse JAML 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 conjugate are sequentially added to microwells pre-coated with a junctional adhesion molecule-like (JAML) 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 junctional adhesion molecule-like (JAML) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Junctional adhesion molecule-like ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Junctional adhesion molecule-like protein (JAML), or AMICA1, is a member of the JAM family of transmembrane proteins. It consists of two extracellular immunoglobulin-like domains, a transmembrane region, and a cytoplasmic tail involved in activation signaling. Known JAML ligands are coxsackievirus and adenovirus receptors, and it has been shown to localize to tight junctions on epithelial cells. | |||||||||||||||||||||||||||||||||
| 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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4.2 ★★★★★
Based on 20 reviews
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Product Reviews
★★★★★ 4
A still timely writing for all pastors
Format: Paperback
Although first published 25 years ago, Peterson addresses a matter that is still timely, the need for all pastors to commit themselves to prayer, reading scripture and spiritual direction. These, Peterson asserts, are an essential foundation for all other aspects of ministry. Peterson's premise, that many pastors have abdicated their primary calling to these three practices, is still very relavent to the current state of pastoral ministry. Its content will be useful for new and veteran pastors alike.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 14, 2013
★★★★★ 5
Transformative if you are willing.
Format: Kindle
The triangle of pastoral ministry
- Scripture - Prayer - Spiritual Direction is reckoned as sever lack in our pastoral lives. Peterson is a needed sage in the realm of modern evangelicalism.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 13, 2024
★★★★★ 5
A Personally Challengiing Read
Don’t expect Rosaria Butterfield to coddle sin. She acknowledges the Biblical truth that we are at war with it. It is evil. Lies are rampant with it. The Bible’s solution is to kill it, nailing it to the cross of Christ. If we don’t, it will do great and lasting harm. That is the background premise of this book.
This book is for believers in Jesus Christ, especially those who identify as Evangelicals. Others may benefit by reading its pages, but I suspect that most of them will not be able to persevere through it because today’s culture is just not willing to sit through this much proclamation of Bible truth. They will react, not respond. Believers will be convicted, encouraged, and edified.
Butterfield’s style is plain, but today’s culture requires her to address and to use terminology that may be challenging to many readers. It was for me. Add a level of abstraction for some challenging spiritual truths. Add the fact that she is an intellectual dealing with cultural and theological subjects. It is mostly college level reading, except for the story parts.
Butterfield has credentials. A professor of English at Syracuse University, she was ten years in the Lesbian community with a mate. She was found by Christ, began to grow in Him, was discipled by faithful church women, met a wonderful man, was married to him--a Presbyterian pastor—became a mother, and began to minister to college students from a Christian perspective. She tells some of her story and the stories of others in the pages of the book. It is fascinating reading.
The book is organized around the five lies that Butterfield has chosen to expose, with two to four chapters dealing with each. She acknowledges that there are others that could be exposed as well. The last one, about our culture's attitude toward modesty, was the most surprising to me.
The book also has a Foreword by Kevin DeYoung, a Preface, and an Introduction. Don’t skip them. The Foreword is nothing short of a good sermon that should be shared in every church in America. In the Preface Butterfield spills the beans, revealing exactly where she stands on key topics and to whom she is addressing this volume. The Introduction is engaging and lengthy and pretty much summarizes the rest of the book.
Along the way Butterfield deals with homosexuality, changing gender, male leadership in the home and in church, the bedrock importance of repentance, progressive sanctification, intersectionality, “gay Christianity” (Side A and Side B), feelings and truth, empathy and sympathy, submission of the wife, feminism, inerrancy, envy and biblical contentment, suffering, modesty and exhibitionism, and even worship.
She quotes scripture, sometimes at length, and she gives copious Bible references. She also has footnotes, some of which are as interesting as the text. She names and uses or responds to contemporary authors on the topics at hand.
She gives solid directions for those dealing with contemporary issues. She also has suggestions for dealing with family members beset by these matters including a question-and-answer section. Page 301.
I recommend this book for all Christian leaders and for believers who want to understand contemporary challenges for the church from our modern culture.
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Reviewed in the United States on May 27, 2024
★★★★★ 5
Great information
Great read helps to communicate with others
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 28, 2026
★★★★★ 5
Relevant and serious issues for today’s Christian
This is a no joke book. She takes Christianity seriously and gives her own life story and growth to encourage the reader. Very convicting read.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 2, 2026
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