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Description
Human TTF1 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 ultrasonically disrupted or repeatedly frozen and thawed. Finally, the homogenate is centrifuged at 5000×g for 5-10 minutes and the supernatant is collected for analysis. 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 a capture antibody against Transcription Termination Factor, RNA polymerase I (TTF1). After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by peroxidase (HRP) catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of Transcription Termination Factor, RNA polymerase I (TTF1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Transcription Termination Factor, RNA polymerase I ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | RNA polymerase transcription termination factor I, also known as TTF1, is encoded by the TTF1 gene. This gene encodes a transcription termination factor that is localized to the nucleolus and plays a key role in ribosomal gene transcription. The encoded protein mediates the termination of RNA polymerase I transcription by binding to the Sal box terminator element downstream of the pre-rRNA coding region. Variation in spliced transcripts encoding multiple isoforms has been observed for this gene. | |||||||||||||||||||||||||||||||||
| 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 and other biological fluids |
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4.3 ★★★★★
Based on 424 reviews
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Product Reviews
★★★★★ 5
Superman: The Golden Age: Volume 1 Review
Format: Paperback
If you’re a fan of, or are interested in the Golden Age of comics, this book is for you. This is really the mainstream beginning of superhero comics. Before everything became mired in continuity, there were one-shot stories that were fun, and often dark. I definitely also recommend this for people who want to get into Superman as a character. For the price, the amount of content you get just can’t be beat.
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Reviewed in the United States on January 31, 2020
★★★★★ 5
This is a Superman I can believe in
Format: Paperback
This is the original Superman, the one who made the character a hit. His powers have limits - a fire threatens his life! - and he uses them for the little guy, against social injustice. One of the best stories, from Action #5, has Supes fighting a breaking dam and flood, but mostly he's fighting human crookedness - crooked lobbyists, crooked football coaches, crooked mine owners, crooked taxi rackets. This Superman is a law unto himself, dependent on nothing but his strength and his personal sense of right. He's a lot more like Samson in that way than he's a Christ figure, and the result is stories in which he lightheartedly smashes slums so the government will have to build decent housing for the poor, smashes cars of reckless drivers, smashes an oil well to bankrupt the crooked promoters. Private property means nothing to him. Neither do legal rights. He's not here to fight for law and order, he's here to fight for justice as he sees it. The police? the government? They're feckless at best, and more often they're part of the problem. There's a strong Progressive sensibility here: if institutions don't benefit the people, the people need to take charge and change things. That's the Superman we see here, and it's the Superman I like best - the original Superman with brute vigor, a passion for justice with no subtlety, and no taking himself too seriously. It's not art, but it's what made comic books. And it still stands up.
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Reviewed in the United States on February 16, 2014
★★★★★ 5
Where it all began
Format: Paperback
Superman was a hit almost from day one, selling not only millions of comics but quickly went on to star in radio shows, movie serials, TV shows, cartoons, movies and every other media under the sun.
And it all starts here. This volume reprints the very first Superman stories from 1938 - the Superman chapters from Action Comics 1-13, the New York World's Fair special and Superman #1, some of the rarest and most valuable comic books ever published.
The art is crude but serviceable, but the stories are surprisingly political. Rather than fighting super villains or aliens Superman spends more of his time taking on corrupt businessmen and politicians. In one early story he ends a war in Europe by kidnapping an arms maker and forcing him to fight in the trenches. After his experience he swears never to make weapons again. This is a Superman who takes on the real issues of his time, and while the solutions are simplistic his goals are a lot more impressive than stopping bank robbers or killer robots.
An early super villain, the Ultra Humanite, puts in a appearance but even his plot is centered around labor unrest rather than death rays.
This is a fascinating look into the history of American comics. politics and popular culture. I recommend it to anyone with an interest in those subjects.
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Reviewed in the United States on June 26, 2011
★★★★★ 4
The Menacing Man of Steel
Format: Paperback
This story tracks Superman's first fifteen stories beginning with Action Comic #1 through Action Comics #13 and also includes the New York World's Fair Comics #1 story and a few pages that Superman #1 added to its reprints of the stories in Action Comics #1-#4.
These fourteen stories features Superman as defender of the weak against a variety of foes including munitions dealers who Jerry Siegel charged with starting wars to line their own pockets, heartless mine owners, gangsters, and slum lords.
Superman's tactics were far rougher than they would become as Superman became a little more mild during the 1940s. Superman,like Batman struck fear in the hearts of criminals. Though Batman needed a cool name and a scary costume, all Superman needed to was to keep dropping and catching suspects until they talked.
Superman's rough edge would begin to get out of line. In Action Comics #8, he decided to solve the problem of slums by tearing them down forcing the government to rebuild as they had during recent hurricanes. The police responded by putting a warrant out for him for understandable reasons.
From here, Siegel made Superman even more forceful culminating in Action Comics #11 which sees the Man of Steel declare war on "Reckless Drivers." Declaring war involves forcibly seizing control of a radio station to broadcast a warning and then destroying all the automobiles in the police impound lot, among other very destructive acts. The stories serve as an almost cautionary tale of the danger of someone with unstoppable and no humility. It reflects the brashness of a 23-24 year old writer. Thankfully Superman would grow in the 1940s into a character that inspired by hope than by fear.
However, despite the more menacing Superman in this book, there are some fun stories in here. My Absolute favorite is Action Comics #6 which features an agent pretending to represent Superman and selling merchandising rights for the Man of Steel, which turned out to be prophetic of the merchandising machine Superman would become. Action Comics #7 features another story of Superman helping out somebody whose just in trouble and needs help. Action Comics #13 introduces the Ultra-Humanite, the first real supervillain, though we only get to meet him briefly.
Overall, this is great for adult Superman collectors who want to read all of his stories. For kids, I'd probably recommend Superman in the Forties for a more balanced look at the Man of Steel.
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Reviewed in the United States on October 26, 2013
★★★★★ 5
Remember old times.
Format: Paperback
Old one but in good condition my son really liked.
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Reviewed in the United States on February 11, 2026