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Description
Human ATR-2 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 angiotensin II Receptor 2 (ATR-2) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB converts to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of angiotensin II Receptor 2 (ATR-2) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Angiotensin II Receptor 2 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Angiotensin II receptor 2 (ATR-2) is a protein encoded by the AGTR2 gene. AGTR2 is an integral membrane protein that binds to two different G protein-coupled receptors. This receptor mediates signaling in pulmonary fibrosis and regulates nitric oxide synthase expression in the pulmonary endothelium. AGTR2 has recently been identified as a target for the treatment of lung inflammation in cystic fibrosis (CF). The region on the X chromosome associated with CF lung disease is located 3′ to the noncoding region of the AGTR2 gene. | |||||||||||||||||||||||||||||||||
| 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 | 0.15-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
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4.6 ★★★★★
Based on 9 reviews
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Product Reviews
★★★★★ 5
Best pillow I've ever had
Size: Queen (Pack of 1)
I don't really write reviews but holy buckets, Batman. This pillow is legit. Back, neck, and shoulder pain is nothing new for me; I've had scoliosis since I was a kid and hyperflexibility in my neck since birth leading to frequent muscle spasms that have caused me to miss work.
Guys. This pillow. It is the most comfortable pillow I've ever had. Like, my neck doesn't hurt and I genuinely didn't remember what that felt like. It's simultaneously supportive and soft and it doesn't sink in. The instructions are super clear to understand and the pillow fluffs up no problem if it gets a little flat. This is a game changer for me and I so hope anyone who buys this has the same experience.
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Reviewed in the United States on June 6, 2026
★★★★★ 5
I highly recommend this pillow, it’s great for someone with neck or back problems.
Size: Queen (Pack of 1), Size: Queen (Pack of 1)
I absolutely love this pillow! When it says it’s adjustable it definitely means that. When you first get it, it’s hard to believe that it’s going to be anything but put it in the dryer for a few minutes and it becomes so large. You will definitely have to remove some of the stuffing unless you like a really large fluffy pillow.
The stuffing is high-quality and once it has been put into the dryer it becomes a fibrous type material that can be fluffed over and over and go back to the same size. It seems to be a high-quality cover on the outer portion and is thick and has a slight cooling affect. It has a separate cover on the inner portion covering the foam stuffing.
I ordered this because I have neck problems and it has been super comfortable for me because I’m able to adjust it multiple times depending on my needs for the particular night when I lay down. Having the extra stuffing means that if the pillow does happen to go flat I can always add the extra that I took out when I received it.
It did have a slight odor when it first came, but after putting it in the dryer, it went away immediately. I do think that’s only due to the plastic that it’s shipped in not anything with the pillow.
I think this pillow is well worth it price and I would highly recommend it for anyone that is a side or back sleeper or has any type of neck problems.
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Reviewed in the United States on February 3, 2026
★★★★★ 5
Amazing!
Size: King (Pack of 1)
Such a great pillow I’m buying another one! You can add or subtract stuffing to your liking. It’s really nice quality, quilted and soft. Best pillow ever. Super comfy and great sleep. Pretty firm with all the stuffing. Perfect the way it came with stuffing on the side. My husband loves it and now I want one.
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Reviewed in the United States on May 15, 2026
★★★★★ 5
Totally worth the money
Size: King (Pack of 1), Size: King (Pack of 1)
Right off the bat, the packaging is absolutely adorable and felt like I was buying a premium product. I love that this pillow is adjustable and as a side sleeper, I needed a pillow that is both firm and supportive of my neck. This hits all the boxes. We will be buying more of these for our bed!
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Reviewed in the United States on June 1, 2026
★★★★★ 5
It works! Quality pillow.
Size: Standard (1 Count), Color: Original White (Cooling)
This does actually provide a cooling effect, and it's very comfortable! The accompanying literature is helpful too. I believe this to be a quality pillow for the price.
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Reviewed in the United States on June 2, 2026
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