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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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4.7 ★★★★★
Based on 2450 reviews
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Product Reviews
★★★★★ 5
Heavy duty
Size: For Medium Dog, Color: Green, Size: For Medium Dog, Color: Green
I just unboxed this so I will update if my opinion changes. I bought the medium dog toy, it’s a a great size, perfect for my 110 pound lab and my 60 pound pit terrier mix. It’s heavy duty so I don’t see this being torn apart anytime soon, it has a slight mint scent (mostly rubber). As of right now, I can confidently say that I see myself buying more from this brand. Hopefully it helps with their stinky breath, if not then at least it’s a fun toy for the pups.
Update: after over a month of play I can say it’s still going pretty strong. My dog did manage to damage it a bit but if you have dogs who are rough on their toys, this is a good option. I was also surprised to find out that there is a squeaker in the base of it, the pups have to bite really hard in order to hear it, heavy duty toy.
I will say one of the only downsides is that it doesn’t seem too appealing to my pups. If there are other toys around they’ll go for the other toys but occasionally will pick this. I still recommend it. Has been lasting us awhile (I have 3 pups).
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 17, 2025
★★★★★ 1
Dog ends up eating plastic ends
Size: For Medium Dog, Color: Grassgreen
At first I thought this would be a great toy but in reality the design doesn’t make sense. The little pieces ‘pricks’ of the cactus get chewed off and swallowed by the dog. So you’re essentially feeding them plastic.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 1, 2026
★★★★★ 5
AMAZING for my Doberman
Size: For Medium Dog, Color: Grassgreen, Size: For Medium Dog, Color: Grassgreen
This toy is the absolute BEST! It has out lasted every other toy by a long shot. So far it has lasted 7 months with a very tough chewer. No other toy has lasted more than a week or two aside from this one. I got this the same week I got my Doberman puppy. I am very impressed how well it has lasted so far. It’s still in great shape but is missing some of the “spikes” and has some tooth indents. Squeezer still works well too. It’s one of his favorite toys.
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Reviewed in the United States on December 1, 2025
★★★★★ 5
My Puppy Loves this Indestructible Dragon
Size: Large, Color: Pink, Size: Large, Color: Pink
My puppy (8 month old black lab mix) loves, loves, loves her dragon. She has had it for 2 months now. It is one of her favorites and gets daily rough play time. It's her go-to toy for fetch and tug games, as well as just general mouthy play. The squeaker gave out after about 4 weeks, but she would binge squeak for prolonged periods (kind of like chomping on gum) so it was almost a blessing when it went silent. She still chomps on it, but at least now I don't have to hear it. Other than the fact that it's a little grungy and bedraggled from so much loving, it is still in amazing shape. The seams and fur are all still intact. There are no visible holes, rips, or weak spots. I love Chew Guard Technology.
This dragon replaced her goDog Iguana (from PetSmart) that developed a tear in the fur after about 4 months of heavy use. (It was superficial, just the outer fur layer and not the innards, but we disposed of it before she could eat more fur.) She can be pretty rough on her toys and I can never predict if any given toy will last minutes or months. The goDog stuffies definitely fall into the "months" category, even with rough daily use.
Edit: The dragon lasted about 6 months of intensive, hard play before it developed a small tear in the fur. I patched it a couple times, but once my pup discovers a weak spot, it's the kiss of death for any stuffed item. We had to dispose of the dragon (so sad) to keep her from eating any more of the fur. We replaced the dragon with the goDog Chameleon, which is still going strong. I think we will go back to the dragon for our next toy because it's just so darned cute. We gave the dragon to my niece for her golden retriever puppy who is very chew-happy. That dragon is also doing well. You've just gotta love that chew-guard technology. This is one tough toy.
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Reviewed in the United States on September 26, 2015
★★★★★ 5
Precious toy for big dogs too
Size: Large, Color: Skinny Green
Not only is this toy cute and fun, but material is thick and durable even for my crazy GSP who is a destruction artist. This is, the favorite toy his go to in the toy box.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 15, 2026
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