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Description
Human ABHD5 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: 1. 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: 1. 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. 2. 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. 3. 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. 4. 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. 5. 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. 6. 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 20 ng/mL). Then dilute to the following concentrations: 20 ng/mL, 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, and 0 ng/mL. Serial dilution method: Take 7 EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 20 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 10 ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves directly as a blank well; there is no need to aspirate the liquid from the penultimate tube. See the figure below for details. 3. Preparation of Biotinylated Antibody Working Solution: 15 minutes before use, centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration using universal diluent (e.g., 10µL concentrate + 990µL universal diluent). Prepare immediately before use. 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 capture antibody against abhydrolase domain-containing protein 5 (ABHD5). 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 abhydrolase domain-containing protein 5 (ABHD5) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Abhydrolase domain-containing protein 5 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | αβ-hydrolase domain-containing protein 5 (ABHD5) is an enzyme encoded by the ABHD5 gene. The protein encoded by this gene belongs to a large protein family defined by an α/β-hydrolase fold and contains three sequence motifs corresponding to the catalytic triad within the esterase/lipase/thioesterase subfamily. Unlike other members of this subfamily, its putative catalytic triad contains an asparagine rather than a serine residue. Mutations in this gene are associated with Chanarin-Dorfman syndrome, a glyceride storage disorder characterized by impaired long-chain fatty acid oxidation. | |||||||||||||||||||||||||||||||||
| 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 | 0.312-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.2 ★★★★★
Based on 1371 reviews
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Product Reviews
★★★★★ 5
Our first babies favorite and longest lasting ball yet and he is fixing to be 7.
Style: Ball, Size: Medium (Pack of 2), Style: Ball, Size: Medium (Pack of 2)
Toughest ball we have found yet. Our doberman loves Chuckit! Balls especially the crunch one. He spreads all the balls and toys we have bought but these lasted the longest. The crunch and the glow in the dark are usually his pick but he has several different choices.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 24, 2026
★★★★★ 5
This is a great ball for high energy dogs.
Color: Blue
I have a Belgian Malinois, so you can imagine the Maligator jaws. Just opened this today and let her play with it indoors. She LOVES IT. It wore her out and it’s a toy she can really stay focused on without needing me to throw or roll for her. It moves on its own! I supervised her play. I was hoping it was too big to get under the couch, but no. It goes right under it. I had to block off the gap so I didn’t have to keep getting it out from under for her. The outter “shell” is very durable! When she had resting moments, she chewed it pretty good - I could hear the material squeaking against her teeth. I was worried she was tearing it up, but nope! Not a mark on it. It’s easy to set up and operate. It needed charging before play. After hours of play, she was able to somehow unscrew the outter shell. This concerned me bc the smaller, inner ball mechanism came out but I saw it right away and was able to put it back together. So, make sure it’s screwed together tightly (which I thought I had done without breaking it) and check it from time to time to make sure it hasn’t loosened. This is a great ball for her incredible high energy drive. It gives her the work and fun play she needs to get tired, which I love. This toy is on the expensive end, but so far it’s worth it. Now we’ll see how long it lasts!
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Reviewed in the United States on October 7, 2025
★★★★★ 5
Love love it
Color: Blue
Best dog toy ever. I have a 1 year old boxer who is full of energy (understatement). This ball has provided more entertainment for us both than any dog toy I’ve ever purchased. Charge holds well and has a very resilient shell. I love it. Worth the price.
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Reviewed in the United States on May 23, 2026
★★★★★ 4
Great ball for limited entertainment. Replaceable outer shell is an amazing option!
Color: Blue, Color: Blue
Bought this for our 45lb, ball-obsessed Dutch shepherd/pit bull dog mix. I had one of these years ago with the hard shell and couldn’t be happier with the new foam shell.
The new foam shell is much quieter and while our dog might put holes in it with his teeth, he doesn’t seem to ever pull pieces off.
For engagement level, he found it VERY interesting at first and was scared to grab it, but once he got over that he started holding it in his mouth even when it shakes and just tries to keep it.
Isn’t the best for fetch or anything as he has trouble letting it go when it is actively moving, but it makes him happy while he has it. We just make sure to monitor him while it is out.
The ability to change the outer shell and just buy a replacement for the shell instead of a whole new toy is a lovely feature that makes me very happy to continue buying from this company.
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Reviewed in the United States on April 19, 2026
★★★★★ 5
Great Interactive Option
Color: Blue
To start, I have been very happy with this purchase. A couple of key points for me are durability and being able to keep my dog's attention. I have had it for almost a year, just a month shy, and she hasn't managed to get through the foam outer shell even though she tries real hard to do so. Let's just say, she's hard to find toys for because of her habit of finding the point of failure where she can pick and gnaw at until the toy is no longer safe for her to have. With this toy, she not only found her match, but it also has her watching, waiting, and even stalking. Only one small issue has recently occurred: the motor has started having an issue where the charge doesn't hold, and it doesn't remain on afterwards. I have reached out to Cheerble directly to see about replacing it since I am still within their 12-month timeframe, and they responded right away.
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Reviewed in the United States on April 22, 2026
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