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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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4.0 ★★★★★
Based on 13 reviews
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Product Reviews
★★★★★ 5
THE favorite toy
Size: 1 Count (Pack of 1), Style: Shark, Size: 1 Count (Pack of 1), Style: Shark
I found this toy used at a thrift store almost 1 year ago. My Shih tzu mix loved it so much till all the seams ripped out and 2 out of 3 squeekers fell out and it was shambles, hardly usable at all. 1 week ago I decided to try my luck to find a similar shark toy for Finnegan. I couldn't believe my eyes when I found THE EXACT same toy!! When I unboxed it, I put the old and new side-by-side and imagine his surprise, it was priceless! Very pleased with this.
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Reviewed in the United States on May 6, 2026
★★★★★ 5
Best Dog Toy Ever!
Size: Small (Pack of 1), Style: Monkey, Size: Small (Pack of 1), Style: Monkey
Unbelievable. See below previous reviews. Today is 3/22/2023 (4th Update): Cavachon is now three years old. THIS MONKEY is STILL played with DAILY. She’s played with and lost interest in dozens of other toys in the past three years, but Monkey is always the go-to. Now only on Monkey #6, which is remarkable considering the daily torture she puts Monkey through. Other toys get destroyed, but Monkey endures. This toy is our dog’s best friend and playmate!
10/9/21 (3rd update): Its rare that any toy (without food inside) keeps the interest of our Cavachon for even 30 minutes… yet day after day, she runs around with Monkey for at least an hour. She’s on her 4th Monkey… and this thing gets mauled, chewed, super-squeaked, fetched, and shook violently EVERY DAY. Durability times ten!
2/21/21 (2nd Update): Crazy pup is now 14 months old, and she plays DAILY with “monkey”! She’s on her third monkey because eventually she pulls off the tail, but each of her first two lasted months.
ORIGINAL REVIEW MAY 2020: There is something really special about this monkey... Our 5-month-old Cavachon LOVES this toy. I recently ordered a second one, in case she wears out the first one. She knows it’s her “monkey” and finds it when we ask her “Where’s monkey?” She fetches it, walks around squeaking it, and she also zooms around the house shaking it furiously by the tail. She’s been playing with, and chewing on all parts of It, DAILY for three months. In the photo, you can compare the brand-new one, still on the cardboard, with the one she’s mauled for three months. Not much difference. I would say this is an extremely durable, and safe, toy.
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Reviewed in the United States on May 15, 2020
★★★★★ 5
Dogs love this toy.
Size: Small (Pack of 1), Style: Monkey
My dogs favorite toy....I have ordered about 6 of these....she loves her monkeys.
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Reviewed in the United States on April 15, 2026
★★★★★ 4
Nice design, pay attention to the size specification
Size: Small (Pack of 1), Style: Monkey
Nice toy with minimal appendages - meaning that body parts are not quickly chewed off. I made the mistake of getting this one along with one for the Large dog. This monkey is intended to be a small dog toy and as soon as I felt it, I knew my English Shepherd of 8 months would probably have it torn up within a day or two.
I really like the design - no ropes or braids or legs, no small pieces for the eyes, etc. The monkey does have a small curly-Q tail but so far it's stayed attached to the monkey. It's durability has surprised me so far.
For years I have had a Jack Russell terrier, which I think might be considered a small/medium dog (at least mine was). This toy would have lasted him weeks or even months, he didn't set out to eat his toys. Now I have an English Shepherd and I forget that she's probably considered a large dog. She rarely chews up anything EXCEPT her toys, but when she gets a new toy, she spends hours "getting to know it" and she's got pretty sharp teeth.
The fabric on this toy is durable, not easily torn. Because there is no stuffing the squeaker is easy for the dog to squeak - and if your dog hasn't had squeaky toys before, that might freak them out just a little, it did my dog, she started off apprehensive of the toy but quickly grew to enjoy it.
Because this toy is hollow and not dense, my dog doesn't chew on it the same way as she did her rope-style toy, and therefore this toy has lasted longer than the KONG Pudge Braidz Pig Dog Toy, Medium/Large toy (which lasted about 10 minutes). Someone else here posted some good pictures of their larger sized dog with this toy, and that dog looks to be about the same size as mine. That person stated the toy didn't last long and showed pictures of how it looked after being dismantled. I expect that to happen to this one since that dog's size is similar to mine, although it's lasted so far more than a day. I guess it partially depends on not just the toy but the dog, too. Because there's no density to this toy, my dog doesn't seem to want to chew it like she does a bone, instead she tosses and wrestles it, occasionally making it squeak, then lies down next to it and takes a nap.
All in all, this is so far, my favorite dog toy brand. It's less dangerous than the Kong toys or the Nylabone toys, and especially less dangerous than the rope style toys. I purchased several other Petstages toys and so far, they have lasted longer than any other brand, including those named. I just should have been more attentive to the recommended dog size, as this toy is clearly not ideal for large dogs. The Petstages Gator toy is better for a large dog. I have also purchased that one and find it much more suitable.
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Reviewed in the United States on February 10, 2015
★★★★★ 3
Could be made much stronger & better
Size: Small (Pack of 1), Style: Monkey
Only 3 stars because the long fur is nasty and dogs chew it and it comes off … choking hazard. I trim it when I get them. The fabric is to easily tear doednt last long they should make this much stronger. I’ve bought 4 of these and about to buy 2 more my dogs love them.
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Reviewed in the United States on January 13, 2026