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Description
ET ELISA KitProduct Specification Usage Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High precision pipette and gun tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37 constant temperature box 4. Distilled water or deionized water Sample processing and requirements: Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4 overnight, then centrifuge at 1000g for 20
Product Specification
| Usage |
Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High-precision pipette and gun tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37℃ constant temperature box 4. Distilled water or deionized water Sample processing and requirements: Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4℃ overnight, then centrifuge at 1000×g for 20 minutes, and take the supernatant, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. Plasma: Collect the specimen using EDTA or heparin as an anticoagulant and centrifuge at 1000 × g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be assayed or stored at -20°C or -80°C, but avoid repeated freezing and thawing. Cell Supernatant: 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., 1 g of tissue sample to 9 mL 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 the tissue cells, the homogenate can be sonicated or repeatedly frozen and thawed. Finally, centrifuge the homogenate at 5000 × g for 5-10 minutes, and the supernatant can be assayed. Preparation before testing: 1. Remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Prepare the gradient working solution of the standard: Add 1 mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, and then gently mix (concentration is 100 pg/mL). Then dilute to the following concentrations: 100 pg/mL, 50 pg/mL, 25 pg/mL, 12.5 pg/mL, 6.25 pg/mL, 3.125 pg/mL, 1.5625 pg/mL, and 0 pg/mL. Serial dilution method: Take 7 EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 100 pg/mL standard working solution into the first EP tube and mix thoroughly to make a 50 pg/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves directly as a blank well; there is no need to aspirate from the penultimate tube. See the figure below for details. 3. Preparation of Biotin-Antibody Working Solution: 15 minutes before use, centrifuge the concentrated Biotin-Antibody at 1000×g for 1 minute. Dilute the 100× concentrated Biotin-Antibody to a 1× working concentration with universal diluent (e.g., 10µL concentrate + 990µL universal diluent). Use the same day. 4. Preparation of Enzyme Conjugate Working Solution: 15 minutes before use, centrifuge the 100x concentrated enzyme conjugate at 1000×g for 1 minute. Dilute the 100x concentrated HRP enzyme conjugate to a 1x working concentration with universal diluent (e.g., 10µL concentrate + 990µL universal diluent). Use the same day. 5. Preparation of 1x Wash Solution: Dispense 10mL of 20x Wash Solution into 190mL 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 them to 4°C. 2. Sample Addition: Add 50 μL of sample or standard of varying concentrations to the corresponding wells. Add 50 μL of universal diluent to the blank wells, followed by 50 μL of Biotin-Antibody Working Solution to each well. Cover with film and incubate at 37°C for 1 hour. (Recommendation: Dilute the sample to be tested at least 1-fold with universal diluent before adding it to the ELISA plate to minimize matrix effects. The sample concentration should be multiplied by the dilution factor when calculating the final concentration. It is recommended to run replicates for all samples and standards.) 3. Plate Wash: Discard the liquid and add 300 μL of 1x Wash Solution to each well. Let stand for 1 minute, then shake off the wash solution and pat dry on absorbent paper. Repeat this process three times (a plate washer can also be used). 4. Enzyme Conjugate Working Solution: Add 100 μL of enzyme conjugate working solution to each well. Cover with film and incubate at 37°C for 30 minutes. 5. Wash: Discard the liquid and wash the plate five times as in step 3. 6. Add 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. 7. Add 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. Calculation of experimental results: Result evaluation: 1. Calculate the average OD value of the standard and sample replicates and subtract the OD value of the blank well as a correction value. Plot a standard curve for 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 competitive enzyme-linked immunosorbent assay (ELISA). Sample, standard, biotin-labeled antibody, and HRP conjugate are sequentially added to microwells pre-coated with a universal endothelin (ET) antigen. After incubation and washing, the assay 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 universal endothelin (ET) content in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | All | |||||||||||||||||||||||||||||||||
| Synonym | Endothelin ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Competition Law | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Endothelin (ET) is a polypeptide with receptors and actions in many organs of the body. Endothelin constricts blood vessels and increases blood pressure. While normally maintained in balance by other mechanisms, when overexpressed, it can lead to hypertension (high blood pressure), heart disease, and potentially other conditions. It is a 21-amino acid vasoconstrictor peptide produced primarily in endothelial cells and plays a key role in vascular homeostasis. Endothelin has been linked to vascular diseases in several organ systems, including the heart, lungs, kidneys, and brain. | |||||||||||||||||||||||||||||||||
| 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 | 1.56-100pg/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, cell supernatant, tissue homogenate, etc. |
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4.1 ★★★★★
Based on 1343 reviews
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Product Reviews
★★★★★ 5
Great quality and folding it in half is a plus. Fits under a seat in car/truck.
Purchased another, as we need one in each car. This is a great ball thrower, especially as it folds in a half. Makes it easy to store under a car/truck seat. Haven’t had any problem with it, great quality!
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Reviewed in the United States on May 4, 2026
★★★★★ 5
Fantastic throwing aid!
Very well made, great quality and doesn't feel cheap at all. It's thick, not flimsy or fragile so you can really swing it to launch a ball. Saves my body that I don't have to bend over to pick up the ball anymore, and saves my shoulder that I don't have to hurl the ball anymore. Another bonus is you don't have to touch a slobbered, soggy ball, or a ball covered in dirt.
The foldable design is fantastic because I walk everywhere with my dog and don't want to carry around a long stick that's awkward to hold. This folds down small enough that I can securely put it in the back pocket of my jeans without worrying about it falling out. The thing I love most about it is there's a elastic loop at the end so when you fold it you can latch it securely. This small detail is outstanding because I can also hang it on the strap of my sling bag and with the loop secured it will not slip off the strap.
This product is so well made and thought out overall.
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Reviewed in the United States on February 14, 2024
★★★★★ 5
Excellent toy for play
My dog would play all day if I threw the ball to him. He loves this even though he's 12. The scooped end allows you to pick up the ball without touching it. After a few runs it's covered in dog slobber. So that's a major plus for me! Also, some health issues affect my walking and bending at times. This helps with the bending. This is a great game to play with your favorite friend!
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Reviewed in the United States on May 5, 2026
★★★★★ 5
Your high-energy dog will love it!
Why did I wait so long to get one of these?!? I am the stereotypical throws-like-a-girl person, and my dog really needs to have an option that forces her to run far and fast. This is it. I can get that ball half a soccer field away at a minimum if I want. The ball is super bouncy, which adds an element of fun to her attempts to tame it and bring it back. Super worth the money (which wasn't much!).
I've never had one of these before, so can't compare to the non-folding version. This one is easy to carry and fits in a drawer for storrage. No way you can fit it in a pocket, especially if you have a phone in one and dog treats in the other.
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Reviewed in the United States on April 14, 2026
★★★★★ 5
Great usability
I prefer this version over the non-foldable stick, as this one I can fit into my bag and carry it out easily. Great size and has a locking feature to prevent it from folding up when throwing. Highly recommend, my dogs love fetch and it is hard to chew if they get their hands on it.
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Reviewed in the United States on September 13, 2025
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