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Description
Human CD40L 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 Cluster of Differentiation 40 Ligand (CD40L) 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 Cluster of Differentiation 40 Ligand (CD40L) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Cluster Of Differentiation 40 Ligand ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | CD40L is a protein primarily expressed on activated T cells and a member of the TNF superfamily. It binds to CD40 (protein) on antigen-presenting cells (APCs), leading to a variety of effects depending on the target cell type. CD40L has three binding partners: CD40, α5β1 integrin, and αIIbβ3. On TFH cells, it promotes B cell maturation and function by binding to CD40 on the surface of B cells, thereby facilitating cell-to-cell communication. Defects in this gene result in an inability to undergo immunoglobulin class switching and are associated with hyper-IgM syndrome. | |||||||||||||||||||||||||||||||||
| 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 | 0.15-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.6 ★★★★★
Based on 1752 reviews
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Product Reviews
★★★★★ 5
Summer wedding kids suit
Color: Navy, Size: 10 Years, Color: Navy, Size: 10 Years
Bought this suit for my son in the 10 youth size and honestly was pleasantly surprised by the quality for the price. The navy color looks sharp and dressy without being too stiff or uncomfortable. The material has a little stretch, which helped it fit nicely and made it comfortable for him to move around in.
The adjustable waist was a huge plus, especially for growing kids. It fit true to size overall, though the blazer sleeves were slightly long on my son. The pants had a nice slim look without being too tight. We paired it with white sneakers for a more modern look and it came out adorable.
This would be perfect for weddings, graduations, concerts, communion, school events, or family photos. Overall, very happy with the purchase and would definitely buy again.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 18, 2026
★★★★★ 5
Very Modern Handsome Suit
Color: Grey, Size: 7 Years, Color: Grey, Size: 7 Years
This is an incredibly sharp looking suit for the little guy. Great price and features, too. I made a mistake in sizing up instead of down and first one was too sloppy on him and would need alterations. He is thin and very thin in the waist. There is a lot of adjustment with buttons in waistband and coat hides the bunching. This shirt and coat will easily take a fuller body than slim I believe. He loves posing in it and feels very classy. Best purchase for a formal suit that will last longer than most kids clothes size wise. Felt bad about returning but careful folding and packing got it good as new.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 20, 2026
★★★★★ 5
Quality
Color: Black, Size: 7 Years, Color: Black, Size: 7 Years
Excellent quality, My son looks incredible
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 5, 2026
★★★★★ 4
Sharp Looking Kids Suit With One Missing Piece
Color: Navy, Size: 7 Years, Color: Navy, Size: 7 Years
I ordered this suit for my 6yo son in a size 7 so he would have a little room to grow and get more wear out of it, and sizing is perfect. He did not need it for any specific occasion, he just wanted a suit because apparently kindergarteners sometimes wake up and decide they are businessmen now.
Let me start with the positives because there are a lot of them. This is a very well made suit. The jacket, pants, and dress shirt are all surprisingly nice quality. The fabric does not feel cheap or flimsy at all. The jacket has good structure, the buttons are well attached, the seams are clean, and all the little details like the collar and pockets look great. It honestly looks like a proper little formal suit.
The pants are nicely made as well and pair perfectly with the jacket. The white dress shirt underneath is also good quality and completes the whole look. Overall, it is a really sharp outfit and would absolutely work for special occasions like weddings, recitals, formal events, church, or family photos.
Now for the reason I am deducting one star. The listing specifically shows and mentions that this set includes a bow tie. That was actually the main reason I chose this particular suit over others. Some options only had a standard tie or no tie at all, but my son specifically wanted a bow tie.
Unfortunately, the bow tie was not included with mine.
That was definitely disappointing because it was one of the main selling points for me when choosing this set. Now I have to go find a separate bow tie that matches, which I was hoping to avoid.
That said, I do want to be clear that this issue does not take away from the actual quality of the suit itself. The suit is genuinely very well made and looks fantastic on him. If your child needs a suit for a wedding, formal event, recital, or something similar, this would absolutely work.
My son mostly just wanted a suit because he thinks it is cool, so now he has one and can show up to school looking like the smallest CEO in kindergarten.
Overall, this is a very nice quality suit set. I just wish the bow tie that was advertised had actually been included.
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Reviewed in the United States on March 10, 2026
★★★★★ 5
Good for the price
Color: Black, Size: 10 Years
Great suit for the price and it includes the shirt which It was a great surprise
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Reviewed in the United States on May 27, 2026