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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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4.3 ★★★★★
Based on 20 reviews
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Product Reviews
★★★★★ 4
A Frother that Won't Turn Off? Great Service. Lost Package. Great Hot Chocolate. Worth It!
Color: Black, Size: Z1 Motor + Batteries
I saw this item (or one similar) on an Alton Brown short video. He recommended a frother and I thought "what the heck? If AB recommends a uni-tasker, it must be something good."
The item came in, loaded with batteries and all. I removed the plastic tab that interrupts the power and the unit kicked on. I wasn't pressing anything. It just started and would not stop.
To return the item is not worth the time or money, especially since none of the "free return" options are anywhere close to me.
Called the company and told them what had happened. I spoke with a very nice woman and after I described what happened, her response was not faux sympathy, but "that shouldn't have happened. We'll make it right."
She offered not only to replace the unit, but send a second one free of charge. When I declined the second one, she said "take it for your troubles. Give it to a friend if you want."
The order was sent out. Tracking via USPS was sent.
The package was supposed to be delivered on a Friday, which was great as we were supposed to get cold temps and snow. I was anticipating making frothy hot chocolate.
Suddenly the tracking showed the package being delivered. It was not. You cannot ask the USPS to find a package for 7 days and my local post office doesn't answer the phone.
Disappointed, I started to make dinner and looked out the window and I could see my postal carrier's truck a block away.
I waddled over to the truck and talked to her. She said "I know where it is."
She had delivered it 3 blocks away. Same house number, but different street.
She went and found the package, brought it to me and Bob's your uncle.
That night I made hot chocolate and used the frother.
I was surprised that the long shaft did not vibrate or waiver. It stays true. The speed is fast, but not overly so.
The thing made a difference. It seemed to make the hot chocolate "lighter" and smoother. I threw a piece of chocolate in the mug, waited a little and then frothed again using the frother.
No issues with the frother.
I now had frothy, really chocolate-ly hot chocolate. For kicks and on the advice of a friend, I added a tablespoon of peppermint schnapps and used the frother again.
Best hot chocolate from a mix I ever had.
The frother worked better than advertised.
Woo Hoo!!
Because the first one didn't work, I have to knock a star off of the rating for the product. But the customer service and actual accountability of the women with whom I spoke is refreshing in the world of product support and customer service.
The item is really good and would recommend whole heartedly.
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Reviewed in the United States on January 11, 2026
★★★★★ 5
High Quality and Surprisingly Powerful
Color: Absolute Black, Size: Z1 Motor + Batteries
The Pros:
Unexpected Power: I usually know exactly what to expect with the products I order, but this unit really blew me away. It is surprisingly powerful and handles everything much better than I anticipated.
Premium Batteries Included: It was a great surprise to find that it came with Duracell batteries. Seeing a name-brand battery included really speaks to the overall quality of the product and shows they aren't cutting corners.
Solid Build: From the moment I set it up, the unit felt sturdy and well-constructed. It is rare to be this genuinely pleased with a purchase right out of the box.
The Cons:
Operating Noise: Because the motor is so powerful, it does produce a bit more noise than some weaker units. It isn't a dealbreaker, but it is noticeable during extended use.
Bottom Line:
This is a high-quality product that exceeded my expectations in both performance and presentation. The fact that it comes ready to go with reliable batteries is a huge plus. I am very pleased with this purchase.
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Reviewed in the United States on May 12, 2026
★★★★★ 5
Great little frother!
Color: Absolute Black, Size: Z1 Motor + Batteries
Works great! I use it for my coffee & creamer, and also my electrolyte mix. Great price which includes Duracell batteries and a hand little stand.
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Reviewed in the United States on May 30, 2026
★★★★★ 5
Decent customer service
Color: Absolute Black, Size: Z1 Motor + Batteries
My frother worked well for a time and through several battery changes. Then the blending stick/wand fell off and completely detached from the handheld part/motor. I contacted customer service and they originally tried to get me to upgrade via a sales tactic and pay the difference for a 'premium model'. I said no thank you and did not ask for anything else. They contacted me back and offered to send me two of the ones I had so that I would have a back up - but without the batteries or stand. I did appreciate their prompt responses, just less gimmicks would have been nice.
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Reviewed in the United States on April 9, 2026
★★★★★ 5
awesome company
Color: Black, Size: Z1 Motor + Batteries
I ordered one of the frothers and it stopped working in about 1 month. The company was so pleasant to work with and immediately sent me a new one to replace the one I had. They offered me an option of a rechargeable one with a little more cost or the battery powered free. I chose the battery powered and received it quickly. They are an awesome company with great prices and stand behind their products. Nice to see this in a company. Will be buying more items from them in the future!!
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Reviewed in the United States on May 4, 2026