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Description
Rat EERa 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: Gently wash adherent cells with pre-chilled PBS, then trypsinize and collect the cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash the collected cells three times with pre-chilled PBS and resuspend 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. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Collect the supernatant for analysis, or store at -20°C or -80°C, but avoid repeated freezing and thawing. 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 an estrogen-related receptor alpha (EERa) 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 estrogen-related receptor alpha (EERa) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Rat | |||||||||||||||||||||||||||||||||
| Synonym | Rat Estrogen-related receptor alpha ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Estrogen-related receptor α (ERRα), also known as NR3B1 (nuclear receptor subfamily 3, group B, member 1), is a nuclear receptor encoded by the ESRRA (estrogen-related receptor α) gene. It was originally cloned based on DNA sequence homology to estrogen receptor α (ERα, NR3A1). Currently, ERRα is considered an orphan nuclear receptor. It has a wide tissue distribution, but expression is highest in tissues that preferentially use fatty acids as an energy source, such as the kidney, heart, brown adipose tissue, cerebellum, intestine, and skeletal muscle. | |||||||||||||||||||||||||||||||||
| 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, cell culture supernatants, and other biological fluids |
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4.3 ★★★★★
Based on 24 reviews
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Product Reviews
★★★★★ 3
the height is only 16"
Size: coffee table leg
The height is only 16". From the description it appears to be 24" wide and 24" high.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 2, 2026
★★★★★ 4
Good DIY Pipe Legs but Powder Coat Quality Lacks
Size: side table leg, Size: side table leg
These industrial style pipe legs make building a DIY end table easy and the kit saves a trip to the hardware store. The fittings go together normally and assembly is simple. Comes individually wrapped. My main issue is the powder coating looks and feels cheap and rubs off easily. The pipes already had marks from touching each other from unboxing. If you do not mind some wear on the finish over time this is a decent purchase. The metal at the hardware store will be thicker and more durable but will cost more. I’d recommend based on ease of having a kit and it being delivered saving me a trip.
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Reviewed in the United States on December 30, 2025
★★★★★ 5
Stable, attractive, and easy to turn into a table with your choice of top
Size: side table leg
The X-shape of the table legs - constructed with solid metal flanges, T-joints, X-joints and 1-inch diameter pipes - provides a strong and sturdy base for a wood top. The legs have adjustable feet pads to prevent scratches and ensure stability. Assembly of the legs is relatively simple (the instructions are clear) but you will need your own tools to attach a top.
The industrial design is especially well suited to rec room or finished basement use. I am adding a 28" diameter raw edge slab of wood from a recently felled tree as the top and it is very attractive. (Photo unavailable because after I made certain it would work I sent the slab out for drying and finishing.)
If you want a custom table and have a particular piece of wood you want to use, these legs are an excellent choice.
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Reviewed in the United States on March 18, 2026
★★★★★ 5
Very nice kit: Beware it is NOT heavy duty plumbing pipe though!
Size: side table leg
I want to give this kit a high rating, but I need to address the "truth in advertising" bit since there may be an important disconnect between what you think you are getting and what actually comes in the box.
Black plumbing used for shelving or table legs is part of an important niche in interior design. I like the industrial loft look myself and have a variety of glass, steel, and masonry elements in my home. This kit fits that look and provides plenty of "strength" for most domestic uses, like shelves or coffee tables. But they are NOT "industrial pipe" or "maximum durability pipe." Note the phrase "cast iron finish feet," which together with the other claims might lead you to think these are cast plumbing fittings. They are NOT!
The feet and flanges feel like electrical conduit fittings, and the pipe sections are likely aluminum. They might be electrical or they might be multi-use, lightweight pipe.
While all this might lead you to think that you're getting a truly retro, heavy-duty pipe kit made of black plumbing pipe, you are not. But, does it matter? Depends. I find the lighter pipe's smooth finish a nice "tone-down" from the rough plumbing look. All together, this kit provides plenty of strength for most furniture and the rustic pipe look, but with a nicer, lighter finish. To me that is a plus--this is a very nice kit and costs less that if you'd pieced it together from real plumbing parts (I priced it out to compare), and looks retro but cleaner. I knew what to expect because while I got this kit free on Vine, I had purchased a nearly identical kit from another seller last year.
So, I'm giving it 5 stars for what it actually is and for the price, JUST BE AWARE THE NARRATIVE IS MISLEADING. Which is unfortunate because just like some people might want the heavier pipe, I'm betting more would be attracted to the look with a cleaner, lighter feel.
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Reviewed in the United States on December 31, 2025
★★★★★ 5
Strong legs, somewhat industrial but trendy also
Size: side table leg
Good legs, some what utilitarian or industrial, they are strong as they are made out of pipe, and assembly is fairly easy once you get the idea of how it goes together. They are roughly 1.5 feet across maybe a bit less adjustable height on each leg, if you have wood floors you may want to put a pad under each leg as they are full metal pipes. Pretty interesting idea and does not cost much.
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Reviewed in the United States on May 31, 2026