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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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4.0 ★★★★★
Based on 28 reviews
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Product Reviews
★★★★★ 3
Pack's Promise was okay but not great
Format: Kindle
Pack's Promise was okay but not great. I won't recommend it to anyone that I know.
PRO:
* Very likable characters
* Lots of steamy scenes that are written very well
* The spelling and grammar are good
* The punctuation is good with the exception of using hyphens instead of commas. Lots of hyphens. Lots and lots of hyphens.
CON:
* Almost no interactions with any characters outside of Madison and the pack
* Nearly no plot. They meet, get together for a heat, agree to make it permanent, done
* Quite a few typos such as extraneous words, missing words and words out of order
THINGS TO KNOW:
* More steamy scenes than storytelling
* A lot of MM & MMM, some MFMM during heat
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 5, 2023
★★★★★ 5
such a good read
Format: Kindle
Madison, Lucas, Grey and Rian were made for each other!!! First time reading from this author and I’m not disappointed!!! Absolutely love the Love in this book and couldn’t ask for a better OV!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on October 25, 2023
★★★★★ 4
Fluffy and Nice Omegaverse
Format: Kindle
… this would have made 5 stars but for 2 reasons.
A.) there were quite a few typos; misspelled words, missing quotations, “the his” mistakes, and various signs that maybe a proofread would do good.
B.) the writing was quite textbook. Late blooming omega is struggling with her new self, finds a absurdly wealthy pack of alphas, every thing is almost insta-love but she resists, then decides to love herself and let everyone be happy.
Rian was my favourite (obviously the author’s favourite too because he got the most page time) but I wish we could see more of his CEO side? He went to work maybe ONCE the entire time. Gray was supposed to be the “growly one” but he turned out to be puppy dog. Lucas was a genius brainiac doctor - but also super alpha with an aggressive hindbrain with a breeding k*nk?? And then there was no actual “breeding”??
Spice 3/5 - normally omegaverse books are super high on messy smut but this was tamer.
Romance 3/5 - insta-love that was then resisted because of personal hangup’s
Plot 2/5 - weird paced head hopping, showing the same scene from different POV’s that made me feel like it was 2 steps backward, 1 step forward.
Humour 4/5 - there were a dozen lines that genuinely made me chuckle out loud
Would have been five stars but the lack of proofreading and the predictable plot made me unable to get up to ADORED IT level - four stars is still and official ENJOYED IT, y’all. This isn’t a bad rating. The “Club Heat” has intriguing possibilities so I’m going to give the second one a shot.
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Reviewed in the United States on March 31, 2023
★★★★★ 5
ABSOLUTELY A MUST for Omegaverse Girls!!!
I ABSOLUTELY LOVE Jillian West and her books!!! I’m so happy I already bought book two and now I have to buy the others for the Assurance Security series!!
Not gonna lie Val kind of annoyed me at the beginning but she grew on me!! Her men are chef’s kisses!!! Holt annoys me some but I can let it slide. I already bought part two so I’m going to be reading that in between work phone calls!!!! DON’T TELL MY BOSS 😂😂😂😂
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on September 30, 2025
★★★★★ 4
Baby bumps and bodyguards
Format: Kindle
Dark, emotional, and unexpectedly tender, Not Ready is an omegaverse romance that delivers found family feels, fierce protectiveness, and a very pregnant heroine who refuses to break. Vale’s on the run from a stalker, but lands in the arms of three private security alphas, cue the swoony tension, fake marriage twist, and slow-burn heat.
It’s a little gritty, a little soft, and a whole lot addictive. If you love protective alphas, high stakes, and heroines with quiet strength, this one’s a must-read.
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Reviewed in the United States on December 18, 2025
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