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Description
dsDNA ELISA KitProduct Specification Usage Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High precision pipette and gun tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37 constant temperature box 4. Distilled water or deionized water Sample processing and requirements: Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4 overnight, then centrifuge at 1000g for 20
Product Specification
| Usage |
Experimental equipment required for the experiment: 1. Microplate reader (450nm) 2. High-precision pipette and gun tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37℃ constant temperature box 4. Distilled water or deionized water Sample processing and requirements: Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4℃ overnight, then centrifuge at 1000×g for 20 minutes, and take the supernatant, or store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. Plasma: Collect specimens using EDTA or heparin as an anticoagulant and centrifuge at 1000 × g for 15 minutes at 2-8°C within 30 minutes of collection. The supernatant can be assayed or stored at -20°C or -80°C, but avoid repeated freeze-thaw cycles. Cell Lysis Buffer: Gently wash adherent cells with pre-chilled PBS, then trypsinize and harvest by centrifugation at 1000 × g for 5 minutes. Suspension cells can be harvested directly by centrifugation. Wash harvested cells three times with pre-chilled PBS and resuspend in 150-200 μL of PBS per 1 × 10^6 cells (protease inhibitors are recommended; if the concentration is very low, the PBS volume can be reduced appropriately). Disrupt the cells by repeated freeze-thaw cycles or sonication. Centrifuge the extract at 1500 × g for 10 minutes at 2-8°C, and harvest the supernatant for assay. 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 100 ng/mL). Then dilute to the following concentrations: 100 ng/mL, 50 ng/mL, 25 ng/mL, 12.5 ng/mL, 6.25 ng/mL, 3.125 ng/mL, 1.5625 ng/mL, and 0 ng/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each tube. Pipette 500uL of the 100ng/mL standard working solution into the first EP tube and mix thoroughly to make a 50ng/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 Biotin-Antibody Working Solution: Centrifuge the concentrated Biotin-Antibody at 1000×g for 1 minute 15 minutes before use. Dilute the 100× concentrated Biotin-Antibody to a 1× working concentration with universal diluent (e.g., 10uL concentrate + 990uL universal diluent) and use on the same day. 4. Preparation of Enzyme Conjugate Working Solution: 15 minutes before use, centrifuge the 100x concentrated enzyme conjugate at 1000×g for 1 minute. Dilute the 100x concentrated HRP enzyme conjugate to a 1x working concentration with universal diluent (e.g., 10µL concentrate + 990µL universal diluent). Use the same day. 5. Preparation of 1x Wash Solution: Dispense 10mL of 20x Wash Solution into 190mL 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 them to 4°C. 2. Sample Addition: Add 50 μL of sample or standard of varying concentrations to the corresponding wells. Add 50 μL of universal diluent to the blank wells, followed by 50 μL of Biotin-Antibody Working Solution to each well. Cover with film and incubate at 37°C for 1 hour. (Recommendation: Dilute the sample to be tested at least 1-fold with universal diluent before adding it to the ELISA plate to minimize matrix effects. The sample concentration should be multiplied by the dilution factor when calculating the final concentration. It is recommended to run replicates for all samples and standards.) 3. Plate Wash: Discard the liquid and add 300 μL of 1x Wash Solution to each well. Let stand for 1 minute, then shake off the wash solution and pat dry on absorbent paper. Repeat this process three times (a plate washer can also be used). 4. Enzyme Conjugate Working Solution: Add 100 μL of enzyme conjugate working solution to each well. Cover with film and incubate at 37°C for 30 minutes. 5. Wash: Discard the liquid and wash the plate five times as in step 3. 6. Add 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. 7. Add 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. Calculation of experimental results: Result evaluation: 1. Calculate the average OD value of the standard and sample replicates and subtract the OD value of the blank well as a correction value. Plot a standard curve for 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 competitive enzyme-linked immunosorbent assay (ELISA). Sample, standard, biotin-labeled antibody, and HRP conjugate are sequentially added to microwells pre-coated with a universal double-stranded DNA (dsDNA) antigen. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by peroxidase (HRP) catalysis and to yellow by acid. The intensity of the color is positively correlated with the amount of universal double-stranded DNA (dsDNA) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | All | |||||||||||||||||||||||||||||||||
| Synonym | Double-Stranded DNA ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Competition Law | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Double-stranded DNA (dsDNA) is a DNA molecule composed of two single-stranded DNA chains linked by base complementarity. Each single strand is linked by alternating deoxyribose sugars and phosphate groups, with each deoxyribose sugar also attached to a base (possibly guanine, cytosine, thymine, or adenine). This single strand forms a planar double helix with the other antiparallel single strand through base pairing (adenine (A) and thymine (T) pair through two hydrogen bonds, and guanine (G) and cytosine (C) pair through three hydrogen bonds). This double helix then spirals into the DNA double helix. | |||||||||||||||||||||||||||||||||
| 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 | 1.56-100 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, cell lysates and other biological fluids |
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4.5 ★★★★★
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Product Reviews
★★★★★ 5
You won't be sorry
Size: 10.5-11.5 Women/9-10 Men, Color: Black
Great padding for comfort and terrific warmth. I bought a pair of Acorns 30 years ago and still have them. Updated to a new color and now feel the benefit of newer padding! I used the sizing listed for shoes and like the roominess for thicker sox. Well made and a joy to put on. (Wisconsin)
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Reviewed in the United States on December 25, 2025
★★★★★ 5
Great Slippers
Size: 12-13 Women/10.5-11.5 Men, Color: Charcoal
Comfy, great price, looks great, fits well, size aligned, great quality. Love these slippers.
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Reviewed in the United States on February 16, 2026
★★★★★ 5
Extremely comfortable, leather sole. Hard to find a style.
Size: 13.5-14.5 Women/12-13 Men, Color: Navy Blue, Size: 13.5-14.5 Women/12-13 Men, Color: Navy Blue
Totally worth the price. This type of style is very hard to find, my husband had another pair a few years ago that he was never able to replace. These fit his wide feet and are very comfortable. The elastic at the ankle is not too tight. So glad I was able to find him a replacement. We bought the blue in a size 12. Love the fact that they are washable.
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Reviewed in the United States on February 12, 2026
★★★★★ 5
Very comfortable Slippers. Great buy!
Size: 12-13 Women/10.5-11.5 Men, Color: Charcoal
I bought these to replace a pair of slippers that I have had for years that are very similar to the slippers I bought, but I had worn a hole in the leather on mine, so I had to find something new and similar. These slippers are worth the money they are very comfortable, and I am sure they will last for years. They are a little tight around the ankle cuff but they are still new and I am sure they will wear in a little more when I start wearing them more.
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Reviewed in the United States on March 8, 2025
★★★★★ 3
Great slippers but disappointing quality.
Size: 7.5-8.5 Women/6-7 Men, Color: Ash, Size: 7.5-8.5 Women/6-7 Men, Color: Ash
Extremely comfortable, attractive slippers but the stitching came loose on both leather trim areas almost immediately. I tried to sew/glue down the stitching (as seen in photo) but it kept unraveling. Disappointing as these slippers are great in every other way (though they do run a bit big...I wear a 9 but ended up with a 7.5-8.5 which fits with a bit of room to spare even with thick socks). I just hope they hold up after my latest attempt to stop the thread from pulling out any more as I do really love wearing them.
Perhaps I just got a bum pair?
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Reviewed in the United States on April 3, 2024