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Description
Human™ 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 freeze-thawed. Finally, centrifuge the homogenate at 5000 x g for 5-10 minutes, and remove the supernatant for testing. For other biological fluids: Centrifuge at 1000 x g for 20 minutes, and remove the supernatant for testing. Pre-Assay 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 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 utilizes 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 a Thrombomodulin/Soluble™ 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 Thrombomodulin/Soluble™ in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Thrombomodulin; Soluble ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Thrombomodulin (sTM) is an integral membrane protein expressed on the surface of endothelial cells that acts as a cofactor for thrombin. It reduces blood coagulation by converting thrombin from a prothrombin to an antithrombin. Thrombomodulin is also expressed on mesothelial cells, monocytes, and a subset of dendritic cells. Thrombomodulin is encoded by the THBD gene. The protein has a molecular mass of 74 kDa and consists of a single chain with six tandem repeats of an EGF-like structure, a serine/threonine-rich spacer, and a transmembrane domain. | |||||||||||||||||||||||||||||||||
| 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 and other biological fluids |
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4.0 ★★★★★
Based on 7 reviews
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Product Reviews
★★★★★ 5
Navarro Has Written a J'Accuse for Our Time, a Bare-Knuckled Indictment of an Evil System
Format: Hardcover
Peter Navarro has written an exceptional book that should be required reading for those who seek to understand the abuses of the Biden presidency. The book is a poignant love story, an account of great sacrifice, and an illustration of the true totalitarian nature of the Left. It is a J'Accuse for our time. Like Zola's work, it is a bare-knuckled indictment of an evil system. Unlike Zola, this system conspired not against one man, but against the President and many others--in fact, well more than half the country. Navarro's account is a contemporary reminder of the abuses Arendt and Solzhenitsyn identified regarding not only the banality, capriciousness, and brutal incompetence of tyranny, but that one does not realize when it starts how horrific it will become. If the Left is not stopped, what they did to Navarro will segue to even greater abuses against more and more people. I Went to Prison So You Won't Have To is a warning to all Americans of what is at stake and hence is a book that Americans must read.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on September 18, 2025
★★★★★ 5
Find Out Who Was the Legal Secretary
Format: Hardcover
I ordered this book after watching Nuremberg with Michael Shannon and Russell Crowe because I wanted to find out who was the woman accompanying Justice Jackson to Germany. Yes, that was not his wife, it was his legal secretary, Elsie Douglas. This book is the only source I could find that goes into any detail about their relationship. What it provides is nowhere near salacious, because it is evident that Jackson took his secrets to the grave. What details it does provide about the evident love triangle that existed in Jackson's life for many years is quite sufficient to explain the events of Nuremberg.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 24, 2026
★★★★★ 4
Well written. Factual sloppiness.
Format: Hardcover
Good insight into Jackson’s approach to the law. Thoroughly researched. Well written. However, Professor White made two egregious factual errors, both of which are an indication of sloppiness on his part, as well as that of his editors and peer reviewers. First, he refers to the World War 2 Chief of Naval Operations as the Secretary of the Navy. Second he states that Los Alamos National Laboratory is in California.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 1, 2026
★★★★★ 4
Judging Jackson
Format: Hardcover
Readers interested in FDR's era, the post-war Nuremberg Trial, and/or such important U.S. Supreme Court events as the Rosenberg spy case and Brown v. Board of Education will enjoy this well-researched book. Illuminating insights as to the characters of his fellow members of the High Court, especially Justices Black and Douglas, are also present.
Of special interest in today's world is Justice Jackson's thinking on the limits of Executive Power in terms of war powers and foreign relations.
I do think non-lawyers will be less interested when the author, a legal scholar, sometime goes into depth exploring aspects of Justice Jackson's thoughts on opaque technical areas, such as diversity jurisdiction under the constitution.
And, I fail to see the need to spend much time on Justice Black's two children, as is done in chapter two "Family Matters." Why do I need to know that his daughter was a mediocre student in college or details about her early love life?
Professor White clearly admires Robert H. Jackson as a fine advocate, brilliant writer, and independent judge, while noting some of his flaws. All in all, a good biography of an important American.
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Reviewed in the United States on November 9, 2025
★★★★★ 5
READ
Format: Hardcover
This is a good book to remind us of the futility of war and the tragedies that occur because of it.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on March 20, 2026
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