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Description
Mouse AdipoR2 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 frozen and thawed. Finally, centrifuge the homogenate at 5000×g for 5-10 minutes, and collect the supernatant for analysis. Cell Lysis Buffer: Adherent cells should be gently washed with pre-chilled PBS, then trypsinized and harvested by centrifugation at 1000×g for 5 minutes. Suspension cells can be harvested directly by centrifugation. Collected cells should be washed three times with pre-chilled PBS and resuspended 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 freezing and thawing or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and collect the supernatant for analysis. 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 10 ng/mL). Then dilute to the following concentrations: 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, 0.15625 ng/mL, and 0 ng/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 10ng/mL standard working solution into the first EP tube and mix thoroughly to make a 5ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves 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 adiponectin receptor 2 (AdipoR2) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by peroxidase (HRP) catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of Adiponectin receptor 2 (AdipoR2) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Adiponectin Receptor 2 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Adiponectin receptor 2 (AdipoR2) is a protein encoded by the ADIPOR2 gene. It is a member of the progesterone and adipoQ receptor (PAQR) family and is also known as PAQR2. Similar to G protein-coupled receptors (GPCRs), AdipoR2 possesses seven transmembrane domains. However, AdipoR2 is oriented on the membrane in the opposite direction to GPCRs (i.e., cytoplasmic N-terminus, extracellular C-terminus), and does not bind to G proteins. AdipoR1 and AdipoR2 act as receptors for globular and full-length adipoins, mediating increased activity of AMPK and PPAR-α ligands, as well as fatty acid oxidation and glucose uptake by adipoins. | |||||||||||||||||||||||||||||||||
| 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 | 0.156-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
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4.5 ★★★★★
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Product Reviews
★★★★★ 5
Potent and Pure — You Can Feel the Difference
Flavor Name: Extra Strong, Size: 3.4 Fl Oz
This black seed oil is definitely strong — you can tell it’s pure and freshly cold-pressed from the first taste. The flavor is bold but clean, exactly what you’d expect from a high-quality oil. I appreciate that it’s ethically sourced and bottled in glass instead of plastic, which helps preserve its freshness.
I’ve noticed a difference in overall energy and digestion after adding it to my daily routine. It’s not the easiest oil to take straight, but it’s worth it for the results. Solid value for the quality and strength.
Black seed oil has become one of my go-to oils for skincare. It absorbs quickly without leaving a greasy film and helps calm irritation, dryness, and redness almost instantly. I like to mix a few drops with my regular moisturizer or apply it directly at night — it leaves my skin soft, hydrated, and with a healthy glow by morning. The natural antioxidants and fatty acids make it great for restoring balance, especially if you deal with uneven tone or occasional breakouts. It’s a strong oil, so a little goes a long way, but the results are worth it — smoother, clearer, and healthier-looking skin.
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Reviewed in the United States on October 12, 2025
★★★★★ 5
Said to have healing qualities - high quality and pure in nice package - but could use a drip spout
Flavor Name: Extra Strong, Size: 3.4 Fl Oz
Super high quality - pure - blackseed oil. If I were to change anything, I would want it in a container that I could control the flow out of. That said - I use this on my knee (which is giving me some trouble these days) - hearing from a co-worker that in another country, this oil used with some prayers, is known for healing. I can't vouch for that - but I do use it as a matter of hoping the oil might get into my joints and help my knee hurt a little less some days. The packaging is great, and I love the bottle. I would just encourage them to put a 'drip spout' on the bottle - because if you know anything about oil, it finds it's way down the sides of the bottle if you aren't careful. Easy clean up since it's a natural substance though. It does have an interesting 'gun oil' smell - to me. It's not bad in any way. And it also seems to absorb quickly into my skin. It is on the brown-ish side, and if you rub your skin against material before it dries - well just remember - it's 'oil'...and may discolor what you touch.
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Reviewed in the United States on May 16, 2025
★★★★★ 5
Great quality!
Flavor Name: Extra Strong, Size: 3.4 Fl Oz
Strong and excellent quality. Best taken with food
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Reviewed in the United States on April 14, 2026
★★★★★ 5
Best quality I've found on Amazon yet
Flavor Name: Extra Strong, Size: 3.4 Fl Oz
I've tried several different brands of black seed oil on here and so far this one seems like the best quality yet. It makes me feel really great when I take it regularly. I try to find one's with 2% tq or higher and this one has it. I see reviews where people won't take black seed oil bc of the taste and that it upsets their stomach and I concur on both, however, there are ways around both. To take, I put a teaspoon under my tongue and take a swig of water after to wash it down, then a few more swigs to get it all down and no taste left in the mouth. I never taste it that way. You also should take it after a meal, not on an empty stomach. It makes me very queasy and burpy if I don't eat first. So I make sure to take it after breakfast and no stomach issues at all! Highly recommend.
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Reviewed in the United States on December 20, 2024
★★★★★ 4
🤠
Flavor Name: Mild, Size: 3.4 Fl Oz
Good oil liking the unknown benefits yet has got to be right!
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Reviewed in the United States on January 15, 2026
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