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Description
Human ATGL 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 freeze-thawed. Finally, centrifuge the homogenate at 5000×g for 5-10 minutes and remove the supernatant for analysis. Cell culture supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, but avoid repeated freeze-thaw cycles. Pre-Assay Preparation: 1. Remove the reagent 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 stand for 15 minutes to completely dissolve, then gently mix (concentration 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 seven EP tubes and add 500uL of universal diluent to each tube. Pipette 500uL of the 20ng/mL standard working solution into the first EP tube and mix thoroughly to make a 10ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube is used 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 adipose triglyceride lipase (ATGL) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of adipose triglyceride lipase (ATGL) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human adipose triglyceride lipase ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Adipose triglyceride lipase, also known as protease domain-containing protein 2 and ATGL, is an enzyme encoded by the PNPLA2 gene. ATGL catalyzes the first reaction in lipolysis, in which triacylglycerols are hydrolyzed to diacylglycerols. ATGL has a very high substrate specificity for triacylglycerols and contains a catalytic dyad that utilizes a serine-aspartate nucleotide. | |||||||||||||||||||||||||||||||||
| 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 | Tissue homogenate, cell culture supernatant |
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4.9 ★★★★★
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Product Reviews
★★★★★ 5
Stunning Automatic Watch With TopTier Durability and Precision
Color: Grey
“I got this Tissot PRX for my cousin’s birthday and bro fell in LOVE instantly.
This thing keeps time like a beast, never slows down, never lags. Automatic movement, so no batteries, no drama.
Durability? Stupid good. You can hit it, bump it, live life rough, it still looks box fresh.
Size wise it was almost perfect. I just had to remove two links, nothing big.
It’s light, functional, comfortable, doesn’t weigh you down, doesn’t get in the way but more importantly I can say this watch is in everyone’s eyesight when we wear it. Cause of course I had to get another one!
Basically. I gave him a very nice piece as a gift, and now he’s walking around like he owns Switzerland.
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Reviewed in the United States on December 11, 2025
★★★★★ 5
Great watch!!!!
Color: Grey
Excellent quality, one of the best, elegant.
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Reviewed in the United States on May 6, 2026
★★★★★ 5
Excellent value for an automatic watch, keeps VERY good time.
Color: Grey, Color: Grey
I've only had the watch for a day but wanted to leave a quick review on it to share a couple of notes.
So far it keeps VERY good time, better than any of the automatics I own (including a Rolex Submariner, Tag Heuer Aquaracer, Tag Heuer Formula One, and several other sub $1k watches). I set it to an atomic time app on my phone yesterday and 24 hours later it's still SPOT on - hasn't gained/lost even a 1/10 of a second. I was shocked to see it's kept PERFECT time for the last day, I'm really blown away so far. I've worn it the entire time, we'll see how well it keeps time while being stored in a winder.
This picture shown is the watch with a 22mm leather band on it, fit's without any issues (just need to squeeze in the leather a tad). I doubt it would work with a 22mm metal bracelet, but leather works great. Just FYI since the 21mm lug width is kind of odd.
The face isn't quite a lustrious as the images make it look, but it's still a great-looking watch. For a sub $1000 automatic you can't go wrong, this is a fantastic watch at a great price. It'll be my daily driver for some time I'm sure. If I have any issues in the future I'll update the review.
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Reviewed in the United States on March 22, 2021
★★★★★ 5
It looks really cool and striking.
Color: Grey, Color: Grey
Really cool and beautiful watch. Although my specific unit runs 5 seconds fast every day—at first it’s annoying, but eventually you stop caring. 🤣
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Reviewed in the United States on April 16, 2026
★★★★★ 5
A Beautiful Timepiece at an Awesome Price!!
Color: Grey, Color: Grey
I have lusted after this watch for four months. I first tried on a similar model (stainless steel band with a green face) in a local jewelry shop (Spitz Jewlers, Walnut Creek, CA, USA). It was "love at first sight," even if I didn't love the white lines on the face from 12-6 and 3-9. At the time, the MSRP was $775.
The MSRP has since increased to $795. But I bought this beauty on Amazon.com for only $524.99; after applying the cash-back from my Prime credit card, this watch costs only $498.75. This was "the offer that I could not refuse," to quote from "The Godfather."
Accuracy could be better: after only a few days, it is now 7 seconds behind my phone (to which I set my new watch). But I don't really care about that all that much. This is why I rated "accuracy" at 4 stars, but my overall review is 5 stars.
My video shows the date changing suddenly at 11:59:59pm, just how I want it; in another photo, you can see that my Seiko changes the date more slowly, which I do not like.
This watch is absolutely phenomenally beautiful. If you're on the fence over whether to make this purchase, then I would say "go for it!" You will not find a prettier, more accurate watch at a lower price in ANY store in the entire USA.
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Reviewed in the United States on April 28, 2023