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Description
Human CYFRA21-1 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: 1. 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: 1. 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. 2. Plasma: Collect the specimen using EDTA or heparin as an anticoagulant. Centrifuge the specimen 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 freezing and thawing. 3. 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 the tissue and mince it. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1 g of tissue sample to 9 mL 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 the supernatant can be assayed. 4. Cell Lysis Buffer: Gently wash adherent cells with ice-cold PBS, then trypsinize and collect cells by centrifugation at 1000×g for 5 minutes. Suspension cells can be collected directly by centrifugation. Wash collected cells three times with ice-cold PBS and resuspend 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 freeze-thaw cycles or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and remove the supernatant for analysis. 5. Cell Culture Supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, avoiding repeated freeze-thaw cycles. 6. 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 Cytokeratin Fragment Antigen 21-1 (CYFRA21-1) 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 Cytokeratin Fragment Antigen 21-1 (CYFRA21-1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Cytokeratin Fragment Antigen 21-1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Cytokeratin 19 fragment antigen 21-1 (CYFRA21-1), located on the 36 kDa fragment of cytokeratin 19, is the most specific tumor marker for NSCLC. It is commonly associated with epithelial cell cancers, including NSCLC, and is often associated with the SQLC type. Because cytokeratins are structural proteins of keratin-containing intermediate filaments found in epithelial cells, their degradation produces soluble fragments that can be measured in the blood of lung cancer patients as tumor markers. | |||||||||||||||||||||||||||||||||
| 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 | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.1 ★★★★★
Based on 14 reviews
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Product Reviews
★★★★★ 5
Great Value for a Comfortable Home Office Chair
Color: Black, Size: Pack of 1, Color: Black, Size: Pack of 1
I’ve been using this chair for a couple of days now, and honestly, it’s been a solid upgrade from my old one. I work from home most days, so comfort matters a lot—and this chair delivers better than I expected for the price.
The first thing I noticed was the lumbar support. It’s actually adjustable and makes a noticeable difference, especially if you tend to sit for long stretches. My lower back used to get sore after a few hours, but that hasn’t really been an issue since switching to this chair. The mesh back is also a nice touch—it keeps things breathable, which I didn’t think I’d care about until I had it.
Assembly was pretty straightforward. Took me maybe 20–25 minutes without any frustration. Everything lined up well, and the instructions were clear enough.
In terms of comfort, the seat is firm but not hard. It doesn’t feel like one of those super plush chairs, but I actually prefer that because it feels more supportive over time. The reclining feature is smooth, and I like that you can lean back a bit when you need a break.
Overall, I’d say this is a great value for a home office setup. If you want something ergonomic and comfortable without spending a fortune, this chair is definitely worth considering.
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Reviewed in the United States on April 30, 2026
★★★★★ 5
Excellent Office Chair
Color: Black+white, Size: Pack of 1
I just received my TRALT Office Chair. First of all, I was impressed with the packaging of this item. When I opened the box each piece of the chair was wrapped in plastic or bubble wrap. Everything in the box arrived safely and without damage. Secondly, the instructions to assemble the chair were very each to follow. I was impressed with the Allen wrench which had a handle attached to it. This made screwing in the screws very easy. I was able to assemble the chair without any difficulty. Lastly, the look of the chair and the comfort is exceptional. I ordered the black/white chair which looks amazing. The comfort of the chair is really excellent. My back is supported by the lumbar seat and having the armrests being able to go up when not needed is wonderful. The quality of the chair is definitely worth the price.
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Reviewed in the United States on May 9, 2026
★★★★★ 4
a good choice
Color: Black, Size: Pack of 1, Color: Black, Size: Pack of 1
Assembled chair only to find that the lift cylinder was faulty. Would have preferred to be able to just get a replacement cylinder but calling China didn't seem like a viable option. Guessing which one & buying it online would involve paying for a good portion of what the chair cost. Customer service could only offer a replacement or refund. We're disassembling (so much fun?!) to send it back. The chair itself was very comfortable, assembled fairly easily and instructions pretty clear. Again, if we could only have asked the seller for the replacement part......**Update**...Or if the instructions that seemed so clear were clearer!, as we started to disassemble, we decided to see what would happen if the tension knob (see photo) was loosened. IT ALLOWED THE LIFT CYLINDER TO WORK. Not at all what the instructions say. At any rate, we're keeping the chair.
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Reviewed in the United States on June 4, 2026
★★★★★ 5
This chair is amazing
Color: Sky Blue, Size: Pack of 1, Color: Sky Blue, Size: Pack of 1
⭐⭐⭐⭐⭐
Absolutely love this chair! It was super easy to assemble—I had it set up in no time with clear instructions and minimal effort. The comfort level is fantastic; the seat cushion is just right and the back support makes it great for longer sitting sessions.
The color is beautiful in person—clean, modern, and adds a fresh pop to the room without being overwhelming. I also really like the smooth wheels and sturdy build; it feels stable and well-made.
Overall, this is a great combination of style, comfort, and value. Highly recommend!
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Reviewed in the United States on May 4, 2026
★★★★★ 5
Excellent Ergonomic Chair
Color: Black, Size: Pack of 1
I am incredibly impressed with this chair. The setup was a breeze and took me less than 20 minutes from start to finish. It is exceptionally comfortable, offering fantastic ergonomic support that keeps my back pain-free during long hours of sitting. The materials feel premium and highly durable. Plus, the included 5-year warranty is incredibly helpful and provides great peace of mind. I highly recommend it to anyone looking for a reliable workspace upgrade!
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Reviewed in the United States on May 25, 2026