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Description
Human TP53BP1 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 frozen and thawed. Finally, centrifuge the homogenate at 5000×g for 5-10 minutes, and collect the supernatant for analysis. Cell Lysis Buffer: Adherent cells should be gently washed with pre-chilled PBS, then trypsinized and harvested by centrifugation at 1000×g for 5 minutes. Suspension cells can be harvested directly by centrifugation. Collected cells should be washed three times with pre-chilled PBS and resuspended in 150-200 μL of PBS per 1×10^6 cells (it is recommended to add protease inhibitors to the PBS; if the cell count is very low, reduce the PBS volume appropriately). Disrupt the cells by repeated freezing and thawing or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and collect the supernatant 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 20 ng/mL). Then dilute to the following concentrations: 20 ng/mL, 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, and 0 ng/mL. Serial dilution method: Take 7 EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 20 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 10 ng/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 the liquid from the penultimate tube. See the figure below for details. 3. Preparation of Biotinylated Antibody Working Solution: 15 minutes before use, centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration using universal diluent (e.g., 10µL concentrate + 990µL universal diluent). Prepare immediately before use. 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 Tumor Protein p53 Binding Protein 1 (TP53BP1). After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase and to yellow by acid. The intensity of the color is positively correlated with the amount of Tumor Protein p53 Binding Protein 1 (TP53BP1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Tumor Protein p53 Binding Protein 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Tumor protein p53 binding protein 1 (TP53BP1), also known as P53 binding protein 1 or 53BP1, is a protein encoded by the TP53BP1 gene. This protein functions in DNA, selects double-strand break repair pathways, promotes non-homologous end joining (NHEJ), and restricts homologous recombination. It plays multiple roles in the DNA damage response, including promoting checkpoint signaling after DNA damage, acting as a scaffold to recruit DNA damage response proteins to damaged chromatin, and promoting the NHEJ pathway by restricting end resection after double-strand breaks. Diseases associated with it include microcephaly and chorioretinopathy, an autosomal recessive form of microcephaly and chorioretinopathy. Pathways involved include cell cycle checkpoints and the ATM signaling network in development and disease. | |||||||||||||||||||||||||||||||||
| 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 | 0.31-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
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4.3 ★★★★★
Based on 8 reviews
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Product Reviews
★★★★★ 5
Indestructible
Style: Fetch Balls (Pack of 2), Size: 3 inch
These balls are indestructible! We have a lab and a lab mix and they CANNOT damage these balls. They love them for fetch and to just chew on them (slightly squishy, but NO damage). Best toys we have bought for them.
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Reviewed in the United States on April 19, 2026
★★★★★ 5
Great trial set. Just be careful throwing the orange one!
Style: Assorted Balls (Pack of 3), Size: 2.5 inch
Blue isn't that easy to see in the yard
White charges with bright light/UV and is truly a game changer for night-time fetch. Both dogs clearly favor it.
Orange is easy to see in the grass at day
They're 3 different weights as well, so double trial. I prefer medium for my big dogs, though light performs well too
The orange is HEAVY and unyielding. Do NOT throw it with a ball thrower. It could damage your house or knock your dog out. Seems tough though.
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Reviewed in the United States on January 16, 2026
★★★★★ 3
Not as good as chuck it anymore
Style: Fetch Balls (Pack of 10), Size: 2.5 inch
I have bought these before and the where just like the chuck it balls. These are not those balls. The are lighter and my dog has already cause damage to one
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Reviewed in the United States on March 16, 2026
★★★★★ 5
A Totally Unique and Lasting Ball Toy Which Captures My Older and Previously Lethargic Dog’s Attention
Size: 3.5"
At about the time of my dog’s 8th birthday, I felt discouraged when I noticed a change in his playing habits and he seemed bored with most of his former favorite playthings. At a younger age, he played for hours until he exhausted himself, and then after a deep and lengthy nap he would resume playing with his toys.
His indoor playing time had decreased even though he was in perfect health. He does not have any arthritis, and he does not have any aches or pain, he was just mostly bored with his toys and after about 10 minutes of playing, he would walk away, stretch out and lie down. He seemed to have lost his interest in chasing after a ball, and I was getting desperate to find something new and completely different to end his daily lethargic existence.
These Gnawsome squeak and light balls have re-captured his interest as something completely different that he had never experienced with any of his other toys. Currently, he has several of these squeak and light balls and in two sizes, the medium 3.5” and the larger 4.5”. There are a variety of colors with different color flashing lights. Both the medium and the extra-large size are suitable for larger breed dogs like my Old English sheepdog. The large dimension guarantees safety that he will not accidentally swallow and choke, because they are too big to slide down his throat.
What makes this squeak ball unique is the addition of a flashing light, as this is something different that he had never experienced with any of his toys.
As he mouths the ball, a light inside the ball starts to flash with color. When he sees the flickering light he then drops the ball and he stares and watches and perks up his ears. When the flickering light stops he bats the ball around, grabs it in his mouth and tries to squeak and flash the light again. It’s obvious that the flashing light fascinates him and he enjoys the squeak sound as he mouths this ball. In addition, the soft flexible rubber spikes must feel good against his gums.
In the mornings as I struggle to wake-up and grab a cup of coffee, he is right there nudging my leg with his ball. He drops it at my feet, and starts running down our long hallway in anticipation of the throw and catch game. At night, it’s even better, I’ll turn off the lights and he can easily see the ball to catch it, and the ball lights flicker as it bounces against our hard tile floor.
I started his Gnawsome Squeak and Light Ball collection almost a year ago with both the large and the medium ball sizes. He has two Reds, two Greens, and one Blue scattered around our home because he carries and plays with them from room to room. Each color ball has a different color light. The Red ones light up red, the Green ones flash a green light, and his Blue one flickers with both a red and a blue light. The only color that we don’t have is the yellow, but I’m still hoping to get that color too.
I am impressed that they have all retained both the squeak and their flashing/flickering lights.
These balls are lightweight so they really go a distance when thrown, and the color NEVER comes off on our white woodwork.
This is one of the best dog toys that I have discovered, they are unique, durable and the squeak and flickering lights last through his mouthing, and normal playing habits. My dog loves them and they are his very favorite toys. In addition, and these light strobe balls may even work for a dog whose hearing has declined, because instead of hearing a ‘bounce’, the dog could just follow and chase the flashing light.
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Reviewed in the United States on August 21, 2017
★★★★★ 5
Great Toy!
Size: 3.5"
My dog loves it. The ball is not soft, that's why I chose it for my super chewer dog. The squeak pitch is a medium level. The ball fits comfortably in her mouth. I would say it's an average weight.
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Reviewed in the United States on February 1, 2026
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