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Description
Human CD83 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 specimens using EDTA or heparin as an anticoagulant and centrifuge 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 freeze-thaw cycles. Cell Lysis Buffer: Gently wash adherent cells with pre-chilled PBS, then trypsinize and harvest by centrifugation at 1000 × g for 5 minutes. Suspension cells can be harvested directly by centrifugation. Wash harvested cells three times with pre-chilled PBS and resuspend in 150-200 μL of PBS per 1 × 10^6 cells (protease inhibitors are recommended; if the concentration is very low, the PBS volume can be reduced 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 harvest the supernatant for assay. 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 200 pg/mL). Then dilute to the following concentrations: 200 pg/mL, 100 pg/mL, 50 pg/mL, 25 pg/mL, 12.5 pg/mL, 6.25 pg/mL, 3.125 pg/mL, and 0 pg/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 200pg/mL standard working solution into the first EP tube and mix thoroughly to make a 100pg/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 83 (CD83) 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 Cluster of Differentiation 83 (CD83) 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 83 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | CD83 is a protein encoded by the CD83 gene. The membrane-bound form of CD83 consists of an extracellular V-type immunoglobulin-like domain, a transmembrane domain, and a cytoplasmic signaling tail. The free, soluble form consists solely of the immunoglobulin-like domain. Membrane-bound CD83 is predicted to form a trimer. Soluble CD83 can assemble into a 12-membered complex. The CD83 gene is located on chromosome 6p23 in humans and chromosome 13 in mice. The upstream 261-bp promoter contains five NF-κB and three interferon regulatory factor binding sites, reflecting the involvement of CD83 in inflammation. A binding site for the aryl hydrocarbon receptor is also present in a downstream 185-bp enhancer sequence located within the second intron and may indicate that transcription is negatively regulated by microbial metabolites produced in the gut. | |||||||||||||||||||||||||||||||||
| 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 | 3.12-200 pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, cell lysates and other biological fluids |
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4.2 ★★★★★
Based on 5 reviews
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Product Reviews
★★★★★ 1
Don’t buy
Size: 28'' X 34''
Mine arrived broken and scratched I think I received one that someone had returned. It also doesn’t hold pads that well my 4 lb teacup Yorkie is able to pull the pad off of the pad holder without even ripping the pad.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 26, 2026
★★★★★ 5
Excellent!
Size: 22'' X 22''
Highly recommend this pad holder. Does an excellent job keeping the pad secure and in place. No more shredded pads since. Bonus: no leaks on the floor. It’s so practical and so helpful.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 1, 2026
★★★★★ 3
Pretty Good But Not Perfect!
Size: 28'' X 34''
I have been using two of these frames for my miniature poodle puppy for almost a month now and while they are useful, they could be improved. The size of the frames are great for our miniature size poodle puppy and the Amazon pads that fit the frames absorb well without leaking underneath. The grips on the bottom of the frame keep it in place so it does not slide across floor. The frame hinges in the center so you can easily carry it through doorways. I am in my 60's so it is nice to be able to carry it to my work table in order to change pads while standing up.
My 4 month old 10 pound puppy can make a dinner plate size circle of pee onto the pad first thing in the morning. 4 large pees are easily held by the pee pads before needing to change it but we prefer to change the pads more often for sanitary reasons. Not sure if you have a very large breed dog that these would work for you. Our puppy likes going on these without hesitation so they are comfortable for him to walk around on and turn to find just the right spot!
Compared to other brands I have looked at this is a great value for money and worth a try to help with potty training your pup. Our puppy sleeps in an X-pen inside of our home at night and when we are gone so these help with keeping the pads where we want them. We have two - one inside X-pen and one by door where we are training puppy to go outside.
These frames are nice in appearance and seem like they will last but they are a little awkward to use compared to another brand I purchased which was more durable and easier to change pad. I ended up with these because other brands do not offer such a large size (28 x 34).
Our main complaints about these frames are:
1. The top frame is hinged and a little wobbly to manuever over the edges of the pee pads. Another brand, while a bit smaller, has a solid one piece frame that goes over entire pad in one step. This frame requires a three step process of laying pad down, pulling edges out flat and placing one side of frame down and then repeating on other side. There is usually a bit of adjustment that needs to be done and so requires opening and closing the frame again for adjustments.
2. The Amazon pee pads that fit this frame hold the pee well without leaking but have very thin tissue paper like edges (blue) that do not extend out far enough to make capturing the edges of the pee pad under the frame very easy to maneuver. It would be a game changer if the pads were made at least 1" bigger around length and width so edges are firmly held in place.
3. Our tiny 10 pound poodle pup is able to easily dig out edges of pad and tear it up. Not as bad as a loose pad by any means, but blue edges are exposed at side centers where frame hinges and there is no cover of edges (unlike a one piece frame) and the blue color seems to attract him. He can dig and loosen pad on any side because there is no way to hide and hold down all of the blue edges of pee pad.
I am still using these to potty train and, as I said earlier, these are way better than loose pee pads. I may shop for even larger size pee pads in the future to better lock down all of the edges without so much difficulty.
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Reviewed in the United States on May 5, 2026
★★★★★ 4
Good product for dog training.
Size: 22'' X 22''
Work for the most part. Dogs will chew on it so wipe it down with dawn dish soap. Hard to clean where it clips down.
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Reviewed in the United States on March 13, 2026
★★★★★ 5
Saving My Floors
Size: 28'' X 34'', Size: 28'' X 34''
Omg, what a game changer. My pups like to destroy the pads whenever they get the chance to. With this they still try but never succeed. I will say that it can be a little sturdier as the handles are a little flimsy, but I manage and it does the job. It's very easy to use when I change the pads and I use 2 pads for coverage. I just placed it where I usually place their pads and it fits right in the spot. They are trained to go outside but on rainy days this comes in handy. To save my floors it's definitely worth the money spent.
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Reviewed in the United States on April 26, 2026
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