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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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4.4 ★★★★★
Based on 459 reviews
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Product Reviews
★★★★★ 5
MUST READ BOOK!!
Format: Kindle
I was in a huge reading slump, when one of my favorite authors recommended this book in her readers group (The amazing Marie Mistry 🫶) and I absolutely DEVOURED this book. Now I’m in the club crying about having to wait until December for Term 2!
This book is about a girl named Pandora who has suffered a life that nobody would ever wish to have. This book is about Pandora learning how to live, learning about herself and what she really wants. She has a long journey to find out all of these things, but she took the first steps in this book and it was beautiful to read. Some of the men in this book are our dream men, Hunter and Reed 😍 and then we have the men who need some work and who we all really just want to beat up until they admit what they really feel, Skel, Bram and Dexter. But that cliffy was a killer and I absolutely cannot wait until the next book!!
You have written an absolute dream of a book Lyra and I eagerly await the next installment in this series!!!
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Reviewed in the United States on June 7, 2024
★★★★★ 5
Addicting!
Format: Kindle
I could not stop reading. It was so refreshing to have a series start so completely different than most fated mates/fantasy academy rh I’ve been reading. From the desert scenery to the magic and feeding plus the psychological trauma the characters are there to deal with. Pandora is absolutely adorable and I totally relate to hiding behind my hair. I love that she’s literally the most scary type of demon but it’s not the usual “badass mc” persona (which I do love a badass that can fend for herself and kick ass from the start but it was a nice change of pace). I’m not usually a big fan of bully within the harem but each character has their reasons for their actions and also conflicting feelings about them. I adore Dex and Reed! Complete opposites but their personalities and inner monologues made them instant favs. I can’t wait to see the character growth with the guys and continued strength for Pandora.
The captivating characters and references to the Fate Hallow series added so much depth and now I need another reread while I wait for book 2. The concept of magic and the unique feeding habits of the demon characters were intriguing. I can't wait for the next book to continue this thrilling journey.
In summary, this book is a must-read for fantasy and magic academy rh fans. With its enchanting characters, nods to the Fate Hallow series, and imaginative concepts, it offers an immersive reading experience that hwill leave you craving for more.
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Reviewed in the United States on June 5, 2024
★★★★★ 4
Best academy I've read this year
Format: Kindle
I need a few things when it comes to a first book of a PNR romance series
1-Good world building (which this totally did)
2-An FMC I can root for (oh hell yes, Pandora is someone I can cheer for)
3-Good drama (can you say GROVEL BOYS!)
4-Enough story to make you feel like you really read something with meat (you saw this book is like 600 pages, yeah?)
5-A hook at the end so I want more! (please, Lyra, gimmie more?!? I need more!!)
Be aware this book is a slow burn, but damn do I feel like there'll be some big payoff when it finally happens. Who doesn't like the buildup?
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Reviewed in the United States on June 4, 2024
★★★★★ 5
Pandora’s Pain, Power, and Passion
Format: Kindle
I absolutely love this new world Lyra Winters has created! The spin on a Demon Academy setting was fresh, unique, and completely addictive.
Pandora is a character who immediately captured my heart. Thought to be powerless and enduring years of brutal abuse from her mother, it’s no surprise that her powers emerge at the exact moment she needs them most. After her mother’s death, Pandora discovers her father is none other than Death himself, a soul eater with a dark legacy. Her journey at the academy is anything but easy, filled with challenges tied to her father’s infamous reputation, her barely controlled abilities, and the cruelty of those around her.
Pandora is easy to root for, you feel every ounce of her pain, resilience, and growth. Along the way she meets Reed, a half-human dream demon who’s kind, steady, and the kind of friend everyone wishes they had. There’s also Hunter, a vengeance demon and counselor connected to her father, who adds another intriguing layer to her story. Then there are the bullies: Dexter, a brooding shadow demon; Bram, a chaos demon with a drinking problem and deep hatred for demon nobility; and Skel, a fear demon wrestling with his own darkness. They might hurt her, but they also can’t seem to stay away when she’s in danger, making for some deliciously complicated dynamics.
This book hits so many of my favorite tropes: friends to lovers, enemies to lovers, and of course, the irresistible “who hurt you?” storyline. I devoured it, and I’m already diving straight into book two!
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Reviewed in the United States on September 4, 2025
★★★★★ 5
So good!
Format: Kindle
Oh my goodness! What did I just read? Lyra Winters you have some serious explaining to do! That cliffhanger killed me! I am so happy to be back in the world of Kalista. This is definitely darker than Fates Hollow but oh so good. This is a fated mates reverse harem which I absolutely love.
Pandora had a very hard and rough upbringing. She lives in pain constantly and it makes it hard on her. She struggles with everything because she was kept so isolated and is new to her magic. Pandora gets sent to the Reform Academy and all of them have reasons why they are there. I love how, after everything she has been through, she is still a nice person. She is growing and becoming stronger too. Love her character.
The guys all act like jerks at first but all have a back story that helps understand why even though want to smack them. I'm here for the groveling that I'm sure will come. I love them all. They each bring something for her. They are all drawn to her though.
This is a slow burn book but there will be more books in the series so sure it will build. Man, that cliffhanger was a doozy and need book 2 now!
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Reviewed in the United States on June 3, 2024