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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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4.3 ★★★★★
Based on 332 reviews
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Product Reviews
★★★★★ 5
4 Paws up! My dogs LOVE them
Flavor Name: Variety Packs, Size: 3 Pack, Set name: Large, Flavor Name: Variety Packs, Size: 3 Pack, Set name: Large
I have 2 large dogs who LOVE THESE.
A Great Pyrenees mix who’s a skinny 80lbs and a lab/terrier mix who’s a chunky 50lbs. They both love bones and can REALLY chew through them quickly so I’m really picky about what I buy them.
Here is what is important to me and how I rate and choose bones. These are my comments on this bone set specifically:
Do they break/splinter?
-They don’t splinter, I have seen smaller pieces that have broken off and when the bone gets down to the end (but this is normal with bones) at which point those pieces can be thrown away. I have never had an issue with them choking, being too small or hurting my dogs though and they are aggressive chewers so I could see this happening with some dogs and never happening with others. 4/5 stars for this.
Do they smell/leave stains on the carpet?
-Nope! No odor and they actually leave my dogs breath smelling better and their teeth cleaner! 5/5 stars
Does it upset my dog’s stomach? (My bigger guy can’t tolerate a LOT of things, he vomits often and also gets diarrhea if he eats the “wrong” thing so I have to be very cautious.)
-NOT at all! No upset tummies. This is always very, very important to me. Make sure you have lots of water nearby for your pups though, these will make them very thirsty. 5/5 stars
Do they hurt my dogs tongue, mouth or teeth?
-Nope, no issues with this. They’re able to gnaw into these aggressively without hurting themselves. 5/5 stars
Are they gone in 2 seconds?
-No! I’m always worried the “stuffed” bones will be left in the dust after the stuffing is all gone but my dogs will still keep chewing on these after the stuffing is gone. They still enjoy the bones and they last for quite some time, weeks at minimum. I can also stick some peanut butter or other treats back in the middle after they get the original stuffing all out of them to “reuse” them if it seems like they’re getting a little bored with them. 5/5 stars
Do my dogs ACTUALLY LIKE THEM?
-They love them. 5/5 stars
Price?
-Great and totally affordable. $20 for 3 of them.. so less than $7/bone. This is comparable to a bone from the pet store and these last a lot longer than something you would find there in the “fresh” grocery-store-like-section of the pet stores.
I’m an avid review reader, especially when it comes to items for my fur babies because I want the best for them. I truly love these bones and will keep buying them.
I will continue to buy these!
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Reviewed in the United States on August 26, 2019
★★★★★ 4
⭐️⭐️⭐️⭐️ Finally Found a Long-Lasting Bone That Actually Keeps My Dog Busy
Flavor Name: Variety Packs, Size: 3 Pack, Set name: Large
I’ve tried a lot of filled bones and chew treats over the years, and most fall into two categories: gone in 5 minutes, or upset stomach later. These filled beef shank bones hit the sweet spot.
The bone itself is thick, dense, and real, not a thin, brittle shell. My medium-large dog is an aggressive chewer, and this keeps him occupied for a solid 30–45 minutes working on the filling, then even longer gnawing on the bone afterward.
The filling smells fresh and appetizing (not rancid or overly greasy), and my dog goes crazy for it. We’ve tried the cheese-style and peanut butter–style fillings, and both were a big hit. No crumbly mess, no weird residue, and no digestive issues so far.
What I really appreciate is that these feel safer than many cheaper alternatives. The edges don’t splinter, and the bone wears down gradually instead of cracking.
Why I’ll keep buying these:
• Long-lasting even for power chewers
• Real beef shank bone, not pressed junk
• Filling dogs actually care about
• Helps with boredom and anxiety
• Good value compared to pet store brands
Minor cons:
• Can get a little smelly after heavy chewing
• I supervise and take it away when it gets small (as with any bone)
If you’re tired of wasting money on chews that disappear in minutes, these are a fantastic upgrade. My dog is obsessed, and I finally feel good about what I’m giving him.
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Reviewed in the United States on February 5, 2026
★★★★★ 5
Best bones out there
Flavor Name: Variety Packs, Size: 3 Pack, Set name: Large
These are the best bones I’ve ever bought. I have a lab puppy and as you can imagine most bones don’t last long around here, but this one keeps him entertained for days.
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Reviewed in the United States on June 4, 2026
★★★★★ 5
Happy doggy dancing my ensue
Flavor Name: Variety Packs, Size: 1.26 Pound (Pack of 1), Set name: Small
I haven’t tried them, but the dogs seem to like them. They do that tippy tappy happy dance when these come out.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 26, 2026
★★★★★ 3
Sturdy
Flavor Name: Variety Packs, Size: 3 Pack, Set name: Large, Flavor Name: Variety Packs, Size: 3 Pack, Set name: Large
Ok so I'm very happy that my standard poodle seemed to really enjoy this product. She was lacking and biting it for hours and days. That said she has still yet to make a dent in the bone. It seems to be too hard and too thick for her. I was surprised how heavy it was! It is large enough it is completely safe to leave her in the other room with. No strange odor or transfer of color. I'd say for the cost it isn't worth it as she is only getting filling and not able to eat the bone for some reason. Then again she is entertained for hours..so maybe so. Doesn't seem to bother her sensitive tummies either.
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Reviewed in the United States on May 7, 2026