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Description
Human CD63 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 capture antibody against Tetraspanin 30 Cluster of Differentiation 63 (CD63). 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 Tetraspanin 30 Cluster of Differentiation 63 (CD63) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Tetraspanin 30Cluster of Differentiation 63 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | The CD63 molecule (CD63) is a protein encoded by the CD63 gene. It is primarily associated with the membranes of intracellular vesicles, although cell surface expression can be induced. The protein encoded by this gene is a member of the transmembrane 4 superfamily, also known as the tetramembrane protein family. These members are mostly cell surface proteins characterized by four hydrophobic domains. These proteins mediate signal transduction events and play a role in regulating cell development, activation, growth, and motility. It can serve as a marker for platelet activation. Deficiency of this protein is associated with Hermansky-Pudlak syndrome. In addition, this gene has been implicated in tumor development. Alternative polyadenylation sites have been found in this gene. Alternative splicing results in multiple transcript variants encoding different proteins. | |||||||||||||||||||||||||||||||||
| 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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Good Body Soap
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Reviewed in the United States on May 1, 2026
★★★★★ 5
” Not so long ago we were sure that such an amazing and beautiful reality must be way off in the ...
Format: Paperback
Eclectic Living?
How are you relating right now to Jesus? Did you see the “you” and the “right now?” We’re all different and in different places in our lives, also in the ways we turn to Jesus and trust him. The gospel of Jesus is rich and varied, with so many facets meeting us in our so multiple needs.
We have been thinking about our “union with Christ.” Not so long ago we were sure that such an amazing and beautiful reality must be way off in the future, at the end of the chain of our “way of salvation,” the ordo salutis. When we go to heaven without any remaining sin, then we’ll see how all Jesus has done for us comes together, that was how we used to think. Then John Murray and others began to show us how union isn’t at the far end but at the very beginning of new life in Jesus. That means that our forgiveness/justification and our godly growth/sanctification belong together, both gifts from the Jesus to whom we belong.
For people who know our theological history that could be alarming though. Wasn’t that what the Reformation 500 years ago was all about? Before, people thought that the main thing was to do the best you can, and then it could be maybe that God would answer that with forgiveness and blessing. But how can you tell when you’re doing enough? The more spiritually alert you were, the less sure. Then came Martin Luther and that breakthrough insight: yes the Lord is holy and you aren’t, but Jesus is! It’s his righteousness that he gives you, and now you can be confident that God is on your side, that when things go wrong it isn’t because he’s mad at you, but probably he’s giving you some kind of “fatherly chastisement.”
Isn’t that wonderful, that firm foundation of the Lord’s unfailing love for you! Being totally sure that what we so desperately need right now is right there in the gospel. Isn’t that all we’ll ever need? In some ways it is, but aren’t we still supposed to grow in our godliness? We’re called to love the Lord with all our hearts and our neighbors as ourselves—now that’s not easy, is it? Luther gave us a real start, but we have to add that now we give our hearts to respond to our Lord’s love with whole-hearted life-changing obedience, right? Lutherans tend to want to stick with #1, forgiveness, and we Reformed want the bigger package. But to do that means work, putting together the loving presence of your Jesus and doing those hard things in your life.
That’s the history, but where are we right now? Way back when I became a believer, it was about “what if you were to die tonight?” That is, in the next six hours how much change can you pull off? Not much, so dying tonight was totally about forgiveness. Back then there was also a lot of teaching about the end of the world and the suffering that would happen then. Most believed Jesus would take us out of that before it got too bad. Interesting, but what if we haven’t arrived yet at the end? So our combo of “tonight” and “sometime way off,” wasn’t much for “what if I have to get up tomorrow morning,” which so far is what life is about.
Sure, people became believers, and were decent afterwards. But did the Jesus gospel really have much to do with their lives? Even if they did their thing and read the Bible every day? Friend Rosemarie tells the world that I have “an eclectic fashion statement.” I really like my bright pink shirt and also my Navaho green bola—so I wear them together. If each is great, then together they have to be stunning, right? Well, they do leave people stunned, eclectically.
So here’s a remarkable Biblical doctrine, say justification, and here’s someone struggling with loneliness. Justification has to be the answer, right? Both are important, so don’t they have to fit? Bone up on justification and watch what happens: not much. Loneliness is a lot about not having a clue about relating to people, how does you forgiveness fit that? Eclectic?
John Leonard’s Get Real helps. When you’re getting to know a not-yet-believer, what do you talk about? Here’s John’s profound answer: it depends! It depends on what’s so hard for him, you learn that by Listening! Then you bring a piece of the gospel to him, one of the “many facets of the gospel!” That is, something out of your own hard life and how the Lord has been blessing you through it, from some part of what Jesus has done for you. I don’t believe John tells us how many facets there are, he’s still collecting them.
John is mostly about not-yets. Now comes David Powlison’s How Does Sanctification Work? He’s about “you, yourself and I.” What’s so hard for you right now? Where is that piece of the Bible that helps you understand and go on to live? Look hard for it, don’t be satisfied with eclectic. That’s going to take prayer and trust in Jesus. Jack Miller taught us to have prayer answered right away: Lord, show me my sin. Now add a David one: show me where I am and how Jesus is there for me. Justification may not be it, don’t look for a cure-all, see how the Lord came in the Bible to people with different lives and gave them exactly what they needed. Talk to other believers too.
I’m glad that biblical counseling came along, people and Bible together, Jay! I’m glad that urban ministry also arrived, now we can learn how to think like a missionary by going only a few miles: see how people different from you are blessed through the gospel in ways you never knew; Harvie! Manny! (Underneath all that comes from seeing the culture under the Bible, how it meets people in that mindset or mess or foolishness. Thank you Meredith Kline, Ray Dillard, Dan McCartney and Doug Green).
Can we count on preachers to model that for us: they know where their people are, don’t they? They can’t preach to fit all those needs at once, but wouldn’t it work to show the basic How? We all can do better with giving them feedback, right? Those liberals just about invented Eclectic, they could see some need and make up a story about it, not bothering with the Bible. We have to watch that we’re not doing Eclectic the other way around though, doing the Bible right and mumbling about how it works. That’s why the Lord has raised up those questioning millennials for us, making clear that what we’re doing so far isn’t much help.
As usual I’m better at spotting the problem than giving the answer. But at least I can give you the beginning: read David’s book!
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Reviewed in the United States on June 12, 2017
★★★★★ 5
Great Little Book
Format: Paperback
The title of this book purports to explain to us how sanctification works in the life of a believer. Most people, in American culture at least, will look at this title and expect something of a "how-to" book, filled with bullet points and flowcharts and methods. But Powlison surprises readers by not pinpointing one way of sanctification or one method of growth in holiness but providing instead a broad range of ways God works to grow us into Christlikeness. Powlison is at his best here: succinct, personal, practical and powerful.
This book is written as a counter to the "one way of sanctification" teaching we sometimes hear today. For example, some teach all we need to do to be sanctified is to focus on the gospel, rehearse it to ourselves, remember it. Others teach that sanctification comes through spiritual disciplines. But Powlison says there are a variety of ways God changes us and we need them all at different times.
Powlison's teaching about Truth Unbalanced and Rebalanced in chapter 3 is worth the price of the book alone. God blessed me through this book. Thank you David Powlison for sharing God's sanctifying work in your life with us.
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Reviewed in the United States on June 18, 2017
★★★★★ 4
Well Written
Format: Paperback
A personal account of the author’s suffering paired with other important experiences and Biblical truths. Theologically sound and a quick read. It was not what I was looking for but it was well written, nonetheless.
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Reviewed in the United States on July 18, 2022
★★★★★ 5
Good condition
Format: Paperback
Great item, great content!
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Reviewed in the United States on March 31, 2026
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