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Description
Human TOP3 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 10 ng/mL). Then dilute to the following concentrations: 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, 0.15625 ng/mL, and 0 ng/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 10ng/mL standard working solution into the first EP tube and mix thoroughly to make a 5ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves as a blank well; there is no need to pipette liquid from the penultimate tube. See the figure below for details. 3. Preparation of biotinylated detection antibody working solution: Centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute 15 minutes before use. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration with universal diluent (e.g., 10uL concentrate + 990uL universal diluent). Prepare and use immediately. 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 3-alpha (TOP3) 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 3-alpha (TOP3) 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 3-alpha ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Topoisomerase III (TOP3), also known as DNA topoisomerase 3-alpha (TOP3A), is an enzyme encoded by the TOP3 gene. It controls and alters the topological state of DNA during transcription. This enzyme catalyzes the transient breakage and reconnection of single-stranded DNA, allowing the strands to pass back and forth, thereby reducing the number of supercoiled strands and altering the DNA topology. This enzyme forms a complex with BLM, which plays a role in regulating somatic recombination. It has been shown to interact with the Bloom syndrome protein. | |||||||||||||||||||||||||||||||||
| 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 | 0.156-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
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4.2 ★★★★★
Based on 259 reviews
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Product Reviews
★★★★★ 4
Works and Mounts Nicely
Style: Data/Line Protection - 12 ft
I wanted to get one of these so I could use it to plug in all of my electronics that I use in my office desk (pretty typical). The desk is in the middle of the room though, so it made it all that much more important that I could "hide" this inside of the desk.
It was a little difficult to figure out where to put the screws so they correctly lined up with the holes in the back of the device, but after a couple of tries, got it to work. It does occassionally fall down. It would be really nice if there were additional holes in the back of the item so you could put 4+ screws into the surge protector so it has the ability to hold up all of the heavy plug in's that are on the market today.
The cord being 12' long was GREAT! This was really the reason that I purchased this model. It eliminates the need to plug this item into an extension cord, which was key.
It's been a good item. Nothing special, but how special can surge protectors get? The 12' cord is great, the item is sturdy and I have been very happy.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on September 21, 2012
★★★★★ 5
Great surge protector
Style: Standard - 7 ft
Reliable product
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on December 3, 2025
★★★★★ 5
Really nice product from an equally nice company.
Style: Data/Line Protection - 6 ft
Really nice product from an equally nice company.
Note: I do not have PRIME since Amazon.com does NOT stand behind their BS mark-up service and have become WORSE since they started pushing it so much at delivering non PRIME items.
My experience with Belkin has always been Way Over the Top good. So I recommend any product in general. Their customer service namely and their delivery, packaging is STELLAR.
This product is good in that it is robust, the white does not easily scratch (matte type), and the product seems to be well designed overall. The LED's are not obnoxious at all. The wire is thick, robust, but has enough travel to be useful.
Only small enhancement would be to put the switch up top instead of at the end of the power strip to provide more options. And maybe add a USB port.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 19, 2018
★★★★★ 5
Works Well And Nicely Designed.......... See Photos~
Style: Data/Line Protection - 12 ft, Style: Data/Line Protection - 12 ft
Delivered as pictured and described. I was a bit concerned because of the super low price, thinking maybe it was a discontinued model due to problems but we've had good experiences with Belkin products so I went with this one...
When facing it plugged into the outlet, the cord does lean right as advertised. Length is also correct. True white, not an almond color, so it looks nice with white outlet and carpet where visible.
The switch is effectively placed on the end where the cord comes out so it's harder to accidentally switch off. Also it lights up on top to indicate green for protected or red for not grounded.
Also designed for a phone line on the opposite end but we haven't had use for that where we are using this surge protector. Will update this review should we utilize that feature or have any issues.
Smart design at a great price. Uploading photos so you can get a closer look. Smile. Hope this is helpful!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on August 22, 2015
★★★★★ 5
Must use
Style: Data/Line Protection - 6 ft, Style: Data/Line Protection - 6 ft
I love Belkin products especially when power outages happen.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 7, 2026
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