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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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4.7 ★★★★★
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Product Reviews
★★★★★ 4
Plenty of fun for Gilbert fans!
Format: Hardcover
As in previous books in the series, Gilbert the Goblin complains about the problems he sees with Valentine's Day, what he calls 'mushy malarkey'. Young readers who don't care for the holiday will find much to relate to in the book. Although, Gilbert's complaints seem a tad trifling. Coconut chocolates are 'Blech'. He seems to have a special issue with the cupid's 'perfect', 'beautiful', and 'luxurious' hair. When Gilbert faces off with a group of cupids, he just barely manages to escape their love arrows, or so he thinks until he backs into a cactus in his own yard and believes he has been pricked by one such love arrow. He then becomes a goblin full of love. He loves his spells, his potions, even himself, but not coconut chocolates. He even welcomes his friends over to celebrate before bidding his pet sock slugs a fond goodnight. Good thing the love spell wears off. This funny book follows the pattern of the previous five books, but the addition of adorable cupids as well as Gilbert's beloved cactuses and pet sock slugs provides plenty of freshness as Gilbert once again (reluctantly) joins in the festivities. Plenty of fun for young fans of the series.
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Reviewed in the United States on February 10, 2026
★★★★★ 5
LOVE this book! ❤️
Format: Hardcover
I am a kindergarten teacher and my students LOVE this series! Valentines are the Worst is a new favorite! The illustrations are amazing as always and the story is funny and sweet, perfect for Valentine's Day!
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Reviewed in the United States on January 17, 2026
★★★★★ 5
The best
Format: Hardcover
One of the best in this series! All kids will love it❤️
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Reviewed in the United States on January 17, 2026
★★★★★ 5
Best way to explore Paris
Format: Paperback
This book is awesome! I enjoyed reading it just as much as my kids did! I love the layout and the way that the book assigns the kids to be secret agents, but defers to the case officers (parents) for the final say. We were in Paris for 1 week and although the weather was less than cooperative, we were able to capture our 100 points and award our 4yo and 7yo their secret agent certificates. I love the fact that you can work on this book in any order and do as little or as much as you want at each attraction. The price was awesome and worth every penny. My 7yo particularly enjoyed the challenges at the Musee D'Orsay and the Louvre and it was a great way to keep those visits focused and entertaining for the kids. My 4yo was a little young to stay engaged with every "mission", but the activities were easy enough that she could help (ex. finding a winter scene in the Musee D'Orsay). I loved that this book engaged the kids but didn't detract from any of our visits (we didn't feel like we had to go "out of our way" to find anything). The only disappointment was that they didn't include Versailles and Sainte Chappelle. I have not only recommended this book to all my of friends traveling to Paris (honestly with or without kids), but I have also already purchased the similar books for London, New York, and Washington DC. Can't wait to see what other cities they add!
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Reviewed in the United States on June 28, 2016
★★★★★ 5
Afun way to see Paris as a kid
Format: Paperback
Great way to have kids really look at the sights of Paris. Encourages group or single competition to do each activity. Lots of visual clues.
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Reviewed in the United States on June 1, 2026