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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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4.6 ★★★★★
Based on 1557 reviews
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Product Reviews
★★★★★ 5
Nice elegant suit for any occasion.
Color: Light Blue, Size: 12, Color: Light Blue, Size: 12
The suit set is very nice, the fabric material is durable and soft. Overall stitching is tight with nice surging of the hems and great construction on the shoulders. Buttons and button wholes are well done. I'm going to iron in the creases on the front of the pants because they weren't folded properly for shipping. The material does hold on to wrinkles so don't be afraid of your iron to get a super sharp look.
The color is a soft baby blue that stands out in a crowd of black suits. With the option of an elastic tie or clip on bowtie this is everything you need to make him look presentable for a variety of occasions.
My only complaint about this suit kit is the dress shirt is stiff and thin. I found some loose threads that I don't trust to not get played with. I'll probably trade it out for something with a little more cotton.
As far as cost goes mid range, but definitely worth the cost. I might consider something with a vest if it's a super formal event, but I'm very happy with how this suit comes together.
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Reviewed in the United States on April 18, 2026
★★★★★ 5
Modern luxury with summer fabric that performs above its price!
Color: Beige, Size: 14, Color: Beige, Size: 14
Wow! My son wore it for Easter Sunday and this suit fits the bill! The linen material is spot on--not too heavy and not too light especially since the weather is warming up now. The jacket is quality made--the design is modern executive, the stitches are strong no flaws to note. The pants are tad bit longer but that's okay, build quality is also great. The white shirt is decent and matches well with the rest of the set. The bow tie and tie itself is a great complimentary to the set, though optional to wear. The whole set just has that quality you would really look for in the price range you would want to pay. Overall, best value for for the, wear right outside the bag, maybe a light ironing otherwise it's perfect! I highly recommend it, no complaints here very happy with this suit!
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Reviewed in the United States on April 5, 2026
★★★★★ 4
Heavy linen character, but a beautifully tailored suit!
Color: Beige, Size: 4, Color: Beige, Size: 4
This is a really nice linen suit! The jacket is beautifully made and well-tailored, with a fun speckled texture that has lots of pretty color variation (we ordered the beige). When I placed it side by side with a similar linen suit from another brand, the difference in the linen texture was really noticeable — this had a much stronger linen character that stands out!
It has three useful pockets on the outside of the jacket, which is great for little hands. The only downside is that one of the inside pockets is just for show (faux), which was a bit surprising and somewhat disappointing for my little guy.
The pants have pockets too, but they aren’t very deep — perfect for small treasures, but not much more. The pants are made from a lighter, thinner single-layer linen compared to the fully lined, double-layer jacket, which makes sense for comfort and easy movement.
The pants didn’t come with a front crease ironed in, so you’ll want to press one yourself. The linen did pick up some creases from being folded in the package, but it doesn’t seem quite as wrinkly as some other linen fabrics I’ve tried. Still, like most linen clothes, especially for boys, the pants will need occasional ironing to keep them looking neat and crisp, especially after a day of play and general wear. The structured, lined jacket stays looking smoother and more put-together with less effort.
It also came with a bow tie, a neck tie, and a tuxedo-style white shirt as extras. The ties aren’t great quality — they’d work fine for quick photos but probably won’t hold up to much actual wear. The formal white shirt feels a bit funny paired with the casual linen suit, but I don’t mind at all. We already have white shirts that we prefer, so we’ll use those instead. For us, the extras are really just bonus items we won’t plan on using.
Overall, it’s a handsome little suit that’s perfect for special occasions like weddings, family photos, or holiday events. My kid looked adorable in it! The stronger linen texture on the jacket gives it a premium feel that I really like. Just be ready for a bit of extra care with the pants to keep everything sharp.
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Reviewed in the United States on April 8, 2026
★★★★★ 5
Great suit set
Color: Light Grey, Size: 4
It is incredibly attractive and includes everything a young child needs for a special occasion.
The material is a 75% polyester and 25% linen blend, and the outer fabric feels remarkably smooth. Its unstructured design allows for an attractive, natural drape, and the blend conveniently allows for machine washing at home.
As is typical with linen-based materials, the fabric can be naturally stiff and prone to wrinkling, so I recommend ironing or steaming it before use. The lightweight feel makes it an excellent choice for the spring, summer, and fall months.
Overall, this suit offers exceptional value for the price.
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Reviewed in the United States on April 12, 2026
★★★★★ 5
Summer wedding kids suit
Color: Navy, Size: 10 Years, Color: Navy, Size: 10 Years
Bought this suit for my son in the 10 youth size and honestly was pleasantly surprised by the quality for the price. The navy color looks sharp and dressy without being too stiff or uncomfortable. The material has a little stretch, which helped it fit nicely and made it comfortable for him to move around in.
The adjustable waist was a huge plus, especially for growing kids. It fit true to size overall, though the blazer sleeves were slightly long on my son. The pants had a nice slim look without being too tight. We paired it with white sneakers for a more modern look and it came out adorable.
This would be perfect for weddings, graduations, concerts, communion, school events, or family photos. Overall, very happy with the purchase and would definitely buy again.
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Reviewed in the United States on May 18, 2026