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Description
Human sCD40L 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 culture supernatant: Centrifuge at 1000×g for 20 minutes, take the supernatant for detection, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. 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 utilizes 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 soluble CD40 ligand (sCD40L). 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 soluble CD40 ligand (sCD40L) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Soluble CD40 ligandELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Soluble CD40 ligand (sCD40L) is contained within platelet granules, and its presence in the blood is therefore a marker of platelet activation. By interacting with CD40 present on endothelial and smooth muscle cells, sCD40L may trigger the release of inflammatory mediators, leading to increased activity of matrix metalloproteinases and activation of the coagulation cascade. | |||||||||||||||||||||||||||||||||
| 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 homogenates, cell culture supernatants, and other biological fluids |
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4.8 ★★★★★
Based on 29 reviews
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Product Reviews
★★★★★ 5
Devotional classic for our time
Format: Paperback
“I’ve always borne a hunger for something that seemed as distant as starlight and as wide as the night sky itself. I want the world and what is behind it, the potent beauty it simultaneously reveals and veils. I think we all do…”
If you can feel the intensity of this hunger in your own life and wonder how to acknowledge it in the midst of too-busy lives and a myriad of distractions, you will be inspired to rest within the tension of this by "Reclaiming Quiet”.
Sarah Clarkson has followed her previous study of God’s goodness, This Beautiful Truth, with this soothing expression of our need for quiet in order to experience the extraordinary. Using a similar narrative of personal memoir and spiritual discovery, she describes her own journey towards being satisfied with the quiet of the ordinary. She writes of how the distractions inherent in our world, while ostensibly a quest for the extraordinary, can actually lead us in the opposite direction, and how learning to cultivate quiet is so necessary. “Quiet in this world; perhaps is most often the vessel that bears us out of darkness and into hope."
Each chapter closes with a gentle prayer and an invitation to ponder our own discoveries through thought-provoking questions relating to what we’ve just read.
Reclaiming Quiet is surely destined to be a devotional classic for our time. I read an advance copy but I’ve already pre-purchased copies to give away and I’m planning to reread Reclaiming Quiet as a personal devotional study, but it would also be perfect as a group read/ study.
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Reviewed in the United States on November 5, 2024
★★★★★ 4
Lovely
Format: Paperback
"Because God does not change and his kindness does not end, though the mountains shift and whole worlds crumble, and to actually believe this means to live a different kind of life from one tossed and overturned by news of every new disaster."
Love the Clarkson family's books and stories. Such a wonderful way with words. I disagree with some of the theology but still consider this a good read.
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Reviewed in the United States on December 4, 2024
★★★★★ 5
Attending to the inmost places of our heart
Format: Paperback
Sarah Clarkson’s newest book, “Reclaiming Quiet” is a masterful journey into the heart of what it means to be quiet before the Lord. I was hooked by the foreword and by the end of the first chapter, I was thinking of numerous friends I wanted to share this book with.
We live in a noisy, attention-distracting world and it’s far too easy to get sucked into one activity after another without one thought of being still, of finding the rest and restoration that our mind, body and soul crave.
Sarah’s writing draws the reader in with the loveliest of vignettes from her life as a Vicar’s wife, mother of 4 and author.
Her writing is lyrically gorgeous-each story comes alive by her excellent word choice and vivid descriptions. She shares her struggles with OCD and really gets to the heart of what it means to find quiet in the midst of the busyness of each day.
This is not a how-to or another step-by -step book, but more of a path that will guide the reader into thoughtful pondering of what it means to be still -to make time to sit quietly and commune with our Creator. A favorite quote from her book:
“One of the great gifts that comes to us when we choose to step away from the chorus and listen to the Holy Spirit, is a capacity for conviction and courage. We need to attend in the inmost places of our hearts, where God speaks…We need to listen from the inside.”
Reclaiming Quiet would make an excellent book club selection for a small group or to explore on your own-you won’t be disappointed. I can see this becoming a bestseller.
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Reviewed in the United States on November 5, 2024
★★★★★ 5
American Bullies at Bretton Woods
Format: Hardcover
There, I said it, and I am an American.
I had heard of the conference but never read about it, and certainly had never heard of Harry Dexter White, but this book goes to great length to explain what happened in this important meeting as World War II was drawing to a close and a plan needed to be developed for a new world order regarding the flows of money to facilitate trade and avoid economic disruptions that the world had seen far too much of.
Steil presents more information on John Maynard Keynes than his American antithesis, Harry Dexter White, and for good reason. Keynes was simply one of the most, if not the most, brilliant intellectuals of the 20th century. His theories of economics were evolving through his life, but he is most remembered for his idea that government stimulus could help alleviate a faltering economy when the private sector failed to do the job, and he was opposed as he said to the "gold cage" that for years had been the standard of international finance. He had a biting wit, coupled with a superior intelligence that far outshone his meager appearance (he was ugly, and knew it) but he was cast in the role of a diplomat to present the case for England as the world entered the post war period.
The problem was that England was broke. She had endured two world wars in the space of 30 years and the empire was begging for funds from Washington, and most of her debt to the US from the Great War was still unpaid. She also had an enemy in FDR, who was determined that the imperial preference of England after the war was to be no more. Her crown jewel, India, was pressing for independence and the empire was in the process of unwinding, as was the strength of the British sterling. Keynes pressed to have the new institutions of the World Bank and the IMF located in London, and the Americans under the leadership of White simply said "hell no."
Enter Harry Dexter White. The name is as deceptive as the individual. He was a son of Jewish immigrants, graduating from Harvard late in life, but brilliant in his intellect and determined that America would rule by the strenght of the dollar and Britain was to be no more as a world power. It was interesting to me to see the Treasury Department so powerful over this whole thing. You may think that the Department of State would have more of an influence because these were important global decisions, but their input was minimal. Regardless, White was a Soviet sympathizer and was just in the process of getting raked over the coals when he died early after the war from a heart attack. Keynes also died at the age of 62, not long after the war. The world remember Keynes and White is more of a footnote. I personally did not like White. He reminded me of a Himmler with his rim glasses and nasty litte mustache. As for his boss, Henry Morganthau, Secretary of Treasury, he was little better. His idiotic plan to strip Germany of all industrial capability after the war and turn it into a nation of small farms was leaked to the press and Goebbels made hay of it, likely resulting in many more American casualities toward the end of the war. Just goes to show that FDR used some strange people in his administration. Thank God his selection of generals was far better.
America was brutal toward the British at Bretton Woods. We often think of the English speaking peoples uniting and working together in true harmony to defeat the fascist nations. That is a myth and this book helps bust it. It shows to me how inhuman America was to our British allies, who bore much of the battle of this war alone, with little hope of survival. It is said that when Winston Churchill learned of the attack on Pearl Harbor, he knew that England would win the war and when he retired, he slept like a baby. Little did he know that the selfishness of the U.S. government would put a boot on the neck of England after the war. Churchill once said that the Germans were either at your throat or under your foot. The later part of that pertains to the American response to England toward the end of the war and after.
A good book. Great information, and highly recommended.
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Reviewed in the United States on March 9, 2013
★★★★★ 4
Easy read on Difficult subject
Format: Kindle
This well-documented book explodes the myth of Bretton Woods. The battle between Harry White and John Maynard Keynes turns out to have been contrived.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 30, 2026
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