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Description
Human ADAMTS8 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 uses a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA). Samples, standards, biotin-labeled detection antibodies, and HRP conjugates are sequentially added to microwells pre-coated with ADAMTS8 (ADAMTS8) capture antibodies. 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 ADAMTS8 (ADAMTS8) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human ADAMTS8 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Thrombospondin A disintegrin metallopeptidase 8 (ADAMTS8) is a member of the ADAMTS (thrombospondin motif-containing degrading enzyme and metalloproteinase) family of proteins. The proprotein encoded by this gene undergoes proteolytic processing to produce the mature enzyme, which contains two C-terminal TS groups and disrupts angiogenesis in vivo, making it a novel angiogenesis inhibitor. Several diseases have been mapped near this gene, most notably lung cancer. | |||||||||||||||||||||||||||||||||
| 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 ★★★★★
Based on 26 reviews
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Product Reviews
★★★★★ 5
Must-Have for Dog Owners (Whow have dogs that like to fetch)
Style: 2.5" Ball
Soon as this launcher arrived I went outside with the dogs to try it out. Long story short: This is a great fetch toy. I am able to launch the ball pretty far with little to no effort, and picking the ball back up is as easy as grabbing it with my own hand.
The toy itself is very sturdy, so much so this is the only fetch-toy I'll ever have to buy. Considering how durable it is, I don't think I'll ever need to buy a replacement. I believe Chuck-It is the most popular option, and because this is the first fetch-toy I purchased, I can't really compare it to anything else. A few outlier reviews said the ball was difficult to launch, and I have to disagree. It has been so easy to use I can't possibly see what the problem is. And no, upper body strength is not a factor as I virtually have none. People can get great distance (Half the length of a football field by my eye) in one simple, easy motion. And if other products can launch the ball farther, so what? How far is far enough? Is your dog going to throw a fit because it went 40 meters instead of 50? No. They will not. For anyone upset this toy doesn't launch far enough, ask yourself this: Is the toy for their dog? Or the human who bought it?
The bottom line is this: It feels like it's built to last, it's inexpensive, and it does what it is supposed to do. So long as a toy is "good enough" I don't see the point in splitting hairs and trying to determine which fetch toy is the absolute best.
Saying it's good enough is a bit misleading though. I can't imagine how a dog toy like this could be any better.
It's a must-have for any dog owner. Unfortunately, only one of my pups likes to fetch...
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Reviewed in the United States on January 6, 2020
★★★★★ 5
Works Like a Charm!
Style: 2.5" Ball
WoW!! This thing is pretty cool! It can pick the ball up off the ground - and with little effort - It can throw the ball 3X Higher and Further than I ever could! It wont hurt my elbow or wrist ( way better than before when I was wearing myself out ) I don’t know how I ever got along without it!! It's a must have if your dog loves playing ball. :-)
Update: I've had this For 7 Months Now- it looks and works Great just like the day I bought it- Thumbs up!
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Reviewed in the United States on September 29, 2024
★★★★★ 5
Better Design - Better Materials - No Sticking
Style: 2.5" Ball
Been using Chuck-It launchers and plastic balls for a very long time and they were always a little sticky on giving up the ball. This makes it tricky to throw grounders with precision and can overshoot the fence in the backyard. A couple of years ago, Chuck-It changed the material to a stiffer plastic that made this problem more pronounced. I even tried custom sanding the cup to get just the right grip.
The Fetch-It launcher (using the Chuck-It plastic balls) uses a three-pronged claw with soft rubbery ends that releases perfect every time. The material costs on this new one are noticeably higher while maintaining a somewhat lower price than the brand name original.
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Reviewed in the United States on January 9, 2026
★★★★★ 3
Meh
Style: 2.5" Ball
It "works" but it isn't very good. The grip is so loose on the ball, it lets go early and doesn't thrown very far. A Chuckit is so much better, I can get 3x the distance with a 25" Chuckit. This thing is kind of a waste.
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Reviewed in the United States on May 25, 2026
★★★★★ 5
Great Value Toy
Color: Crinkle Duck (Blue), Size: Large, Color: Crinkle Duck (Blue), Size: Large
This toy has been one of my dog’s absolute favorites. He gets excited every time he sees it and will carry it around the house, toss it in the air, and keep himself entertained for a long time. The shape and texture seem perfect for him, and it quickly became one of his go-to toys over everything else in his basket.
What I really like is that it holds up reasonably well for the price. It’s definitely not indestructible, and after a lot of play, it will sometimes start to rip—usually around the feet first since that’s where my dog tends to grab and chew the most. But honestly, considering how affordable it is, I don’t mind replacing it when needed. It’s inexpensive enough that rebuying feels worth it because he genuinely loves it that much.
I’d rather keep buying a toy he is obsessed with than spend more on something that just sits untouched. It’s fun, cute, and keeps him happy, which is what matters most. Overall, I’d definitely recommend it for dogs who love plush toys and playful chewing.
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Reviewed in the United States on April 28, 2026
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