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Description
Human UBE2L3 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: Gently wash adherent cells with pre-chilled PBS, then trypsinize and harvest cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be harvested directly by centrifugation. Wash the harvested cells three times with pre-chilled 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 freezing and thawing or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and collect the supernatant for analysis. Pre-Assay Preparation: 1. Remove the kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Prepare the gradient working solution of the standard: Add 1 mL of universal diluent to the lyophilized standard. Let stand for 15 minutes to completely dissolve, then gently mix (concentration 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 0.10 ng/mL. Serial dilution method: Take seven EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 10 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 5 ng/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 Ubiquitin Conjugating Enzyme E2L3 (UBE2L3) 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 Ubiquitin Conjugating Enzyme E2L3 (UBE2L3) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Ubiquitin Conjugating Enzyme E2L3 | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Ubiquitin-conjugating enzyme E2C-binding protein E2L3, also known as UBCH7, is encoded by the UBE2L3 gene. As an E2 enzyme, UBE2L3 participates in the ubiquitination of target proteins for degradation. UBE2L3's role in the ubiquitination of NF-κB precursors has led to its involvement in several major autoimmune diseases, including rheumatoid arthritis, celiac disease, Crohn's disease, and systemic lupus erythematosus. | |||||||||||||||||||||||||||||||||
| 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 | 0.156-10ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenate, cell lysate |
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4.6 ★★★★★
Based on 13 reviews
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Product Reviews
★★★★★ 5
Wonderful book
This is the best baby book ever in my opinion. It's very detailed and informative but the way it's set up doesn't make it overwhelming. Also, this book has lots of beautiful, colorful, bright pictures in it. I purchased this book along with Pregnancy Day by Day. I definitely recommend this book!
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Reviewed in the United States on May 21, 2017
★★★★★ 5
Perfect Replacement for Loofahs
Scent: White with Green Edge
I really like these myHomeBody Dual-Textured Exfoliating Washcloths. Where other options only have the roughness, this set has a soft side and a rough side. Having the soft side not only increases how far soap goes (these produce a ton of suds!), but also makes the towels easier to grip and use.
The rough side is the perfect roughness for replacing loofahs, and is great for exfoliating and managing things like keratosis pilaris.
I am so happy with these! I am definitely ordering more.
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Reviewed in the United States on May 2, 2025
★★★★★ 5
My new washcloth every time I shower at home or the gym
Scent: Gray
I initially used an Axe loofa with the scrub pad on one side and a plastic mesh on the other. However, I also wanted a washcloth to clean crevices such as ears, toes, and *other* parts. This product marries the two parts I want (cloth/scrub). The cloth side is soft for cleaning inside the ears, on eyelids, etc., and the scrub side is perfect for the rest of my skin and makes it smooth and healthy-looking. I use a fresh one every time I shower so there is no bacteria growth, and I use it when showering after gym workouts. Also, there's a loop on them so they can hang from the water handle when you're not using it.
I wash them with my regular laundry without damage to my clothes from the scrub side; just let them dry completely before placing them in the hamper so mold doesn't grow. I've had them for 6 months and they still look new, so it's worth the extra $ for that reason. My only suggestion is that they offer to sell them in packs of 5 or 7 at a lower price per washcloth, so it's cheaper to buy a week's set in one purchase.
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Reviewed in the United States on November 23, 2023
★★★★★ 5
If you're partial to mesh loofahs, get these instead!
Scent: Gray
Here I am, a 33 year-old guy and avid death metal fan. Writing reviews for, and getting excited about exfoliating shower cloths. I'm not sure what brought me to this point in life, but I'll attempt to share some wisdom now that I'm here.
Before these cloths entered my life, I was supremely dedicated to showering with traditional mesh loofahs. They had a good "scrub factor" and showering with bare soap bars or normal cotton wash cloths just made me feel like I was pushing soap around without actually getting clean. Well, I eventually got frustrated with having to keep a bunch of them stocked in the bathroom cabinets and the enormous amount of waste they produce. It's recommend to replace traditional mesh shower loofahs every 8 weeks, but uhhh.. who are we kidding? You'd be lucky to use one for half that length of time before the knot loosens up and it all unravels on you.
Enter: these things! These double-sided wash cloths take everything I enjoyed about mesh scrubbers and put it all into a different form-factor. Both the soft side and the exfoliating side make a rich lather with a minimal amount of body wash, which is something you WON'T get with regular wash rags. These cloths are strong, well-made, and mostly importantly they won't unravel into a pile of cheap plastic. Another advantage these have over regular cotton rags is that they're made of a quick-drying material, which makes it a lot harder for bacteria to harbor and grow. A thorough rinse along with a firm wringing to expel the excess water before hanging it up to dry, and you're good to go! One more advantage these have over loofahs and sponges is how much better the cloths are for getting into tighter spaces (I'm specifically referring to cleaning feet and in-between toes, but yes, it'll get THOSE areas nice and clean too) 👌.
At $20 for a 3-pack (on sale for $15 at the time of this review), these cloths are more expensive when compared to the cheap mesh loofahs that you can pick up for $2-3 a piece. But trust me when I tell you that these cloths will far out last even the most premium loofahs, and they clean infinitely better as well.
Give 'em a shot, and maybe one day you'll start getting excited about boring, dumb adult stuff too! 🤘
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Reviewed in the United States on April 4, 2022
★★★★★ 4
I like it just not the soft side
Scent: Gray
Now when I looking for cloths. I was looking for a face one. There was a photo here that showed a up close shot of the texture of the rough side... that's what made me get it. And I'm honestly happy that I did.
I literally just came out the shower. Im air drying while writing this lol. To talk about how it's worth the money getting this. Im surprise they don't have more ratings.
When you look at the video for this product it looks dramatic cause it is. Its meant to be. but when they actually mention details about the product...... its the honest truth.
The rough I love it. I actually feel like I have a new skin after rubbing all the dead skin off my body and used another one to rub my feet (never mix the two lol) so the rough side of the product has 100% 👍🏽 👍🏽 from me.
But the soft side. Ehh honestly it isn't for me if the soft side was a bit more rough. I would love it. But when I used it it felt like I was just rubbing batter on me or something like that.
If they made a glove version of this I would buy 1 for each day of the week lol. If they another cloth that didn't have such a soft side I'll buy that instead.
But honestly this product is amazing and worth the money they're asking form. Which isn't a lot. Believe me you'll feel better after that good scrub down... just be gentle at certain areas and on your neck.
I'm going to buy the actual face one from this company on my next pay check. So worth it
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Reviewed in the United States on January 5, 2022