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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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4.5 ★★★★★
Based on 1925 reviews
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Product Reviews
★★★★★ 5
SUN-sational Sunscreen
Style: SPF 70, Size: 2.5 Fl Oz (Pack of 1)
This sunscreen has become a staple in my daily routine! The Neutrogena Sport Face SPF 70+ goes on smoothly without feeling greasy or heavy, and it doesn’t leave a white cast on my skin. It stays put even when I’m sweating or in the water, which is perfect for workouts and outdoor days. My face feels protected all day, and it hasn’t irritated my skin at all. Definitely worth the price for reliable, strong sun protection!
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Reviewed in the United States on February 12, 2026
★★★★★ 5
Protection is top notch.
Style: SPF 70, Size: 2.5 Fl Oz (Pack of 1)
Can't go wrong with this sunscreen. I found it easy to rub in with no greasy feeling afterwards.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 2, 2026
★★★★★ 5
🚨 Listen up Black and brown-skinned girls, this is for US! APPLICATION is the key!
Size: 1.7 Fl Oz (Pack of 1), Style: SPF 50 Liquid Drops
As the line goes from Love Jones, "Let me break it down, so it can forever and consistently be broke."
It bugs me when a great product gets mediocre reviews when the problem isn't the product, it's the application. I saw another review by a brown woman like me where her face is ghostly white and she claims this isn't for us. The review is still up for you to go find yourself. I almost didn't purchase because of that review but I'm glad I decided to see for myself.
First, physical sunscreens, like this one, are more effective than the chemical ones. Additionally, I am 57 years old and have tried a LOT of chemical facial sunscreen products over the years and even my favorites like the Neutrogena SPF Serum 60 makes my eyes tear. Even if I avoid placing near my eyes, once I'm out and about if the wind blows wrong or I have a little hot flash sweat, my eyes are tearing for the remainder of the day until I get home and relieve myself by washing my face. So there's the backstory.
This sunscreen right here is so good that I did something I almost never do. I recorded a review video without being all "done up". I had to do it on the spot because I'm that impressed. The problems people are having with the severe white cast is because they're applying way too much and rubbing instead of tapping it in. That's the key takeaway from this review.
First, your skin has to be moisturized. I drink approximately 80 ounces of water MINIMUM a day, but my skincare regimen is heavy on the moisture. So in the morning, for instance, I use Hanskin Bubble Pop skin cleanser, the the Hanskin essence, and I also use 2 actives, alpha arbutin and niacinamide. I finish up with Visible Difference Replenishing HydraGel Complex. So by the time I get to the sunscreen step, my face is dewy and ready.
Then, I SHAKE THE SUNSCREEN BOTTLE AGGRESSIVELY for like 15 seconds. There's a little ball inside the bottle to mix it well and the product is thin like nail polish so it needs to be mixed every time you apply.
I drop LITERALLY 3 DROPS OF THE SUNSCREEN DROPS ON THE BACK OF MY HAND. Then I tap just a little bit (see videos) onto a fingertip and begin TAPPING the product onto my face, one area at a time. I admit, yes, it takes a little time, but so? I'm about the long-term health of my skin, which means spending a little extra time when I have to. I'd say to apply it to my entire face takes all of 5 to 7 minutes which is the only step in my morning skincare regimen that takes time. That's not a big deal. When I'm done, there is the very sliiiiightest white cast but it is absolutely minimal. It's not even a bother.
I admit I wouldn't feel confident leaving home with just this sunscreen and no makeup. Brown ladies know what I mean. Even if it's the slightest white cast, you see it in the harsh light of day and office lighting. It's just what it is, titanium and zinc do that to us and there's no perfect way around it if you want the benefits of physical sunscreen.
But here's the thng. I don't wear liquid foundation and most days I just use the Fenty Pro Filt'r Instant Retouch concealer(shade 385) under my eyes and to cover the blemishes on my cheeks. This sunscreen plays nicely with my concealer. But for the video I took for this review, I skipped concealer altogether just so you could see me spots and all.
What I DO have on is just the slightest-teensy dusting of Fenty Pro Filt'r Powder Foundation (shade 385) and that's all I need to conceal any white casting to show you how I'm street-ready with a full face of sunscreen on underneath. That's it. Nothing else needed. I don't look like a ghost and I'm standing right in front of floor to ceiling windows with natural light. No filters, no AI.
Do not try to save time by dropping it directly onto your face and then rubbing it in. It will never work, no matter your skin tone. If you have patience and take a few extra minutes to apply it properly, you can feel confident using this high quality, reasonably priced physical sunscreen.
Lastly, I double cleanse at night with Hanskin PHA oil cleanser first to remove every trace of sunscreen from my face. This stuff is waterproof and if you don't believe me, put a drop on the back of your hand and then hold it under fully running water. It does not move, which is great for warmer weather and girls like me who have hot flashes. I'm not mad about it. I just feel better loosening it up with an oil cleanser before washing it away with my Hanskin Bubble Pop (or whatever other cleanser I'm using for the evening which is sometimes the Hanskin Vitamin C Glow Powder).
My skin has not been irritated, I've seen no additional breakouts and bonus, my actives are working better. It's like night and day from just using the chemical sunscreen I've used for years. No teary eyes, very comfortable to wear and no scent.
If you use it as I've described here, this bottle will last you a minimum of 6 months, likely more. If you're using more than 7 drops for your entire face, you're using too much.
Whew! I've given you all I've got. Now, go forth and be beautiful. Prevention beats correction every time.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on December 6, 2025
★★★★★ 5
Gentle and Reliable for Accutane Skin, Just Not Matte
Size: 1.7 Fl Oz (Pack of 1), Style: SPF 50 Liquid Drops
For the past few weeks, I've been using Cetaphil Sheer Mineral Liquid Sunscreen SPF 50 on my skin while I've been using Accutane and have been putting on this product every morning before I go outside. With Accutane, my skin is very dry and super sensitive but this product doesn't sting or irritate my skin at all. It's nice and gentle on my skin, and I've been getting a good consistent amount of protection from it since my skin burns very easily right now.
The consistency of this sunscreen is very light, almost like lotion, and it goes on easily without tugging on dry skin. The initial application has a little bit of a white cast, but after about one minute, it completely absorbs into the skin. The finish provides a nice soft, hydrated look as opposed to a matte finish. This product does really well with the dryness since most of the products I use tend to make my skin appear more oily.
For daily use, I feel like I have to purchase this product fairly frequently because the bottle runs out rather fast (especially if you apply a good amount). I would recommend this product as a non-irritating option for anyone who is currently on Accutane or has sensitive skin. You can expect this product to have a bit of a dewy finish so you may need to apply again throughout the day if necessary.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 10, 2026
★★★★★ 4
Mineral SPF with very little cast, dewy finish
Size: 1.7 Fl Oz (Pack of 1), Style: SPF 50 Liquid Drops
This is another mineral sunscreen I’ve been trying, and compared to others, it definitely leaves less of a white cast. The formula feels more fluid and lightweight, almost similar to some Japanese-style sunscreens that have a thinner, shake-and-pour consistency. It spreads easily and blends faster than thicker mineral creams.
On my skin, it looks more sheer overall and doesn’t cling heavily to texture. That said, it does leave a slightly greasy or dewy finish that can feel a bit much, especially if you already have combination or oily skin. I personally need a light powder on top if I don’t want to look shiny!
It offers broad-spectrum SPF 50 protection and feels gentle enough for sensitive skin. I haven’t experienced stinging around the eyes, and it layers reasonably well over moisturizer. Because it’s more liquid, I recommend applying in thin layers and allowing it a moment to settle before makeup.
Overall, it’s a solid mineral sunscreen with a more elegant texture and less cast, just be aware of the dewy finish.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 24, 2026