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Description
Human TRAF6 ELISA KitProduct Specification Usage Required experimental equipment: 1. Microplate reader (450nm) 2. High precision pipettes and pipette tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37C incubator 4. Distilled or deionized water Sample preparation and requirements: 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 and
Product Specification
| Usage | Required experimental equipment: 1. Microplate reader (450nm) 2. High-precision pipettes and pipette tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37°C incubator 4. Distilled or deionized water Sample preparation and requirements: 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 and mince the tissue. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1g of tissue sample to 9mL 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 collect the supernatant for analysis. Cell Lysis Buffer: Adherent cells should be gently washed with pre-chilled PBS, then trypsinized and harvested by centrifugation at 1000×g for 5 minutes. Suspension cells can be harvested directly by centrifugation. Collected cells should be washed three times with pre-chilled PBS and resuspended 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 freezing and thawing or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and collect the supernatant for analysis. 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 enzyme conjugate are sequentially added to microwells pre-coated with a TNF Receptor Associated Factor 6 (TRAF6) 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 TNF Receptor Associated Factor 6 (TRAF6) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human TNF Receptor Associated Factor 6 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | TNF receptor-associated factor 6 (TRAF6), also known as RING finger protein 85 (RNF85), is a protein encoded by the TRAF6 gene. The protein encoded by this gene is a member of the TNF receptor-associated factor (TRAF) protein family. TRAF proteins associate with and mediate signal transduction by members of the TNF receptor superfamily. This protein mediates signaling not only from members of the TNF receptor superfamily but also from members of the Toll/IL-1 family. | |||||||||||||||||||||||||||||||||
| 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 | Tissue homogenates, cell lysates, and other biological fluids |
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4.7 ★★★★★
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Product Reviews
★★★★★ 5
Amazing Smart Vacuum — Worth Every Penny!
Color: Black, Size: X60 Max Ultra Complete
This robot vacuum has completely changed how we clean our house. The suction power is incredibly strong, and it handles both vacuuming and mopping very well. The obstacle avoidance is impressive — it moves around furniture smoothly without getting stuck.
The self-emptying and self-cleaning station saves so much time, and the app is easy to use. It’s also much quieter than expected for such a powerful machine. Floors look spotless every day with almost no effort.
Definitely one of the best smart home purchases we’ve made. Highly recommend for busy families and pet owners!
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Reviewed in the United States on May 7, 2026
★★★★★ 5
The Ultimate hands-free cleaning experience
Color: Black, Size: X60 Max Ultra Complete
This robot vacuum is amazing, the 35,000 Pa suction is incredibly powerful, it cleans my floors perfectly. I love the ultra-thin design because it fits easily under all my furniture The self-cleaning and refilling features save me so much time. It is very smart and avoids obstacles well. Highly recommended
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 4, 2026
★★★★★ 5
Convenient and save time
Color: Black, Size: X60 Max Ultra Complete
I have it for about a week now and I am really happy with happy with this purchase so far. This robot vacuum/mop works way better than I expected. It has strong suction and picks up dust, dog hair, and little dirt. The mopping feature is great addition, leaves the floor looking clean and fresh. Setup was pretty easy and the app was simple to use. Great sensor quality and very smart! It moves around furniture good and doesn’t bump like my older robot vacuum cleaner. Overall, very worth it for me. It help me saves time and keep the house clean without much effort.
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Reviewed in the United States on April 29, 2026
★★★★★ 5
Smart, powerful and worth the price 👍
Color: Black, Size: X60 Max Ultra Complete, Color: Black, Size: X60 Max Ultra Complete
This robot vacuum has exceeded my expectations. One of my favorite features is how thin it is it reaches under furniture that my previous robot could never access. The suction is incredibly powerful and does an excellent job picking up pet hair, dust, and debris from both hard floors and rugs. The mopping function is surprisingly effective, especially with the hot-water cleaning that helps tackle dried-on spots and everyday messes. I’ve also noticed how intelligently it moves around the house, avoiding obstacles without constantly getting stuck. The self-cleaning dock takes care of most of the maintenance, which makes the whole experience much more convenient. Setup was straightforward, and the app gives plenty of customization options. Overall, it’s a reliable, hands-off cleaning solution that keeps my floors looking great with very little effort from me.
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Reviewed in the United States on May 31, 2026
★★★★★ 1
Don't toss mop pad holders after a pooptastrophie & take a pic or video for proof
Color: White, Size: X60 Max Ultra Complete
I’ve been using the DREAME X60 Max Ultra Complete Robot Vacuum and Mop for a while now, and it genuinely felt like a major upgrade from previous robot cleaners.
The first thing that stands out is the 35,000Pa suction power—it handles fine cat and dog hair, even on plush carpets. Compared to other models I've had, the cleaning performance is noticeably stronger and more consistent.
The ultra-thin 3.13-inch design is another big win. It reaches under most beds, sofas, and low furniture where dust usually accumulates. It takes some time to program these details into the vacuum.
I was very impressed by the fully automated system:
• Self-emptying dustbin
• Automatic water refilling
• Mop self-cleaning
This significantly reduces manual maintenance—you can go days (or longer) without needing to intervene. Until one Sunday evening, I decided to run the unit in the bathroom I share with my cats' automatic litter box, sight unseen. My cat pooped on the floor right in front of her litter box. Over the past two months, the unit has been good about avoiding wires, my black UGG slippers (the unit thought was an uncleanable mess), and my animals. As I was preparing dinner, I noticed the unit sounded a bit strange and clicky. Cat poo wafted through the air as I saw the unit flinging feces into the air and across the area in a sand-like spray. I thought the unit sucked up the poo, but it was mostly dispersed throughout the bathroom floor, baseboards, and walls. The poo was partially dry when the machine discovered it. By the time I discovered the machine, the mop pads and wheels were caked. I put on some gloves, pulled the mop pads off, tossed them in the trash, and proceeded to clean the machine carefully and gently for the next two hours. The next day, I called customer service, and when I asked for them to send me replacement mop pad holders, they said don't worry about it, we will send a new unit after you provide the info we request in an email. It wasn't until the next day that I got an email back asking for pictures and video of the pootastrophie. I didn't take a picture or video because I trusted that the machine caught it on video and labeled the poo as large particles or something. The next time I logged in, the history and video of Sunday night's ordeal were erased. I'm not sure if there was an update. I received an email today saying they don't have the mop pad holders; instead, they offered to give me more mop pads and dust bags. I can't use the mop function, which is half of the reason I got the unit.
**** Update Friday, May 8, 2026: My claim was escalated after I refused the extra mop pad holders and dustbags nix the mop pad holder. They are following through with replacing the unit and returning the pooptastrophy unit for testing. Will post updates.
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Reviewed in the United States on May 6, 2026
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