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Description
Human TOP2a 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 DNA topoisomerase IIα (TOP2a) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB converts to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of DNA topoisomerase IIα (TOP2a) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human DNA topoisomerase IIα ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Topoisomerase IIa (TOP2a) is an enzyme encoded by the TOP2A gene. It relieves topological stress on DNA during transcription, condensing chromosomes and separating chromatids. It catalyzes the transient breakage and reconnection of the two strands of double-stranded DNA, allowing them to pass through each other. This enzyme exists in two forms and is likely a product of gene duplication events. | |||||||||||||||||||||||||||||||||
| 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 | 0.312-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
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Product Reviews
★★★★★ 5
Feat of diaper engineering
Size: Preemie, Unit Count: 30
I liked these well enough until my preemie had the biggest poo I’ve ever seen. That diaper probably weighed almost as much as he does and his white footie pajamas were untouched. Miracle. Worth every penny. Absolute legend.
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Reviewed in the United States on May 19, 2026
★★★★★ 5
Perfect thank you
Size: Newborn, Unit Count: 144
These baby diapers have been an excellent choice. They are very soft against the skin, which helps prevent irritation or chafing—even for babies with sensitive skin. Their absorbency is excellent, keeping the baby dry for several hours, both day and night. Furthermore, the fit is comfortable and secure, allowing the baby to move freely without any leaks. Without a doubt, they are practical, reliable, and provide peace of mind for parents. Highly recommended!
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Reviewed in the United States on May 3, 2026
★★★★★ 4
Outstanding, top of the line diaper!
Size: Preemie, Unit Count: 30
We normally use Huggies Little Movers with our 3.5 year old (nothing against them, they're also great diapers) but recently decided to give Swaddlers a try, and will not be going back. This is a top-of-the-line diaper and excels in all ways.
We use a size 7 though could *probably* get away with a size 6, and use about 4-5 diapers a day on average (not including his overnight diaper) depending on how much he's had to drink and how often he poops -usually once a day, but could be as many as 3 times, especially if he doesn't go one day.
The first thing you notice about these diapers is how soft they are. I mean, they just feel like they would be comfortable on a child, and since they're in them practically 24/7, that's important. The diaper is mostly white, and the overall designs are more muted than a lot of other diapers, with "Shilo and Friends" -an elephant and duck - on the front and back. I personally wouldn't mind something bolder, I don't think my child cares or even notices. There is a wetness indicator that goes down the length of the diaper, front and back. Nice to have I suppose for a caregiver who maybe doesn't change a lot of diapers, but at this age and stage, it's easy to know and tell. Like all Pampers diapers, they are scented. I happen to love the gentle powder-like scent, but it's a turn-off to some who describe it as too strong, and of course, if your child is sensitive to that, you may want to avoid Pampers diapers.
Inside, the top sheet liner keeps his skin dry, even when he wets heavily -which is usually - and does a good job of keeping poop from sticking to his bottom. The "blowout barrier" in the back also helps contain any messes. In terms of absorbency, I'm pretty amazed at how much they can hold. Being a little older, he tends to stay drier for longer periods and then pee a larger amount at one time, but the diaper almost always holds it in, and leaks are rare.
The one thing I wish that Pampers did do was have wider tapes, like Huggies, at least on the larger sizes, where kids are more active. It hasn't been a problem, so I don't really have a reason for saying that, other than that it seems like it would help with fit, but it certainly won't keep me from buying or using them in the future.
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Reviewed in the United States on May 18, 2026
★★★★★ 5
Tastes great and works very good.
Size: 200 Count (Pack of 1)
I take melatonin often and this has always been the brand and type I have used. It works great for me. Love that it is chewable - easy to take right before bedtime.
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Reviewed in the United States on January 10, 2026
★★★★★ 5
Sound to sleep
Size: 200 Count (Pack of 1)
Love this stuff. Down for sleep in 20 after use. We've tried others. This is the best.
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Reviewed in the United States on October 21, 2025