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Description
Human CCR4 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. 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 utilizes 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 Chemokine CC-Motif Receptor 4 (CCR4) 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 Chemokine CC-Motif Receptor 4 (CCR4) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Chemokine C-C-Motif Receptor 4 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | C-C chemokine receptor 4 (CCR4) is a protein encoded by the CCR4 gene. The protein encoded by this gene belongs to the G protein-coupled receptor family. Chemokines are a group of small, structurally related proteins that regulate cellular trafficking of various white blood cell types. Chemokines also play a fundamental role in the development, homeostasis, and function of the immune system. They influence cells of the central nervous system and endothelial cells involved in angiogenesis, or vascular dilation. CCR4 and CAF-1 form a delipidase complex that collaborates with the release factors eRF3 and PABPC1 to shorten the poly(A) tail of mRNA during translation. | |||||||||||||||||||||||||||||||||
| 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 | Serum, plasma, tissue homogenates and other biological fluids |
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4.3 ★★★★★
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Product Reviews
★★★★★ 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
★★★★★ 5
Comfortable, Sturdy, and Great for Long Hours
Color: Black, Size: Pack of 1
I am very happy with this office chair. It was easy to assemble, feels sturdy, and is very comfortable to sit in for long periods of time. The ergonomic design provides good back support, and the adjustable features make it easy to find the perfect position. The mesh back keeps me cool throughout the day, and the chair rolls smoothly. Overall, it offers great quality for the price and has made working at my desk much more comfortable. I would definitely recommend it to anyone looking for a reliable office chair. ⭐⭐⭐⭐⭐
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Reviewed in the United States on May 30, 2026
★★★★★ 5
45 minutes assembly -- piece of cake. Nice chair.
Color: Black, Style: With Headrest, Color: Black, Style: With Headrest
Before I finally landed on this chair I read about a million articles with buying advice for ergonomic desk chairs, looked for second hand expensive chairs, read so many Amazon reviews. Obviously you can spend a whole lot of money on an ergonomic desk chair (even used ones), but I really didn't want to.
I just bought this today, so the jury is out on long term wear, ,but I can say that the components feel solid, assembly was easy, it is *very* comfortable so far, and -- even more to the point -- VERY adjustable in so many different ways that physical therapists etc recommended in the articles I read. I am 5 foot 5; in order to put the chair at the right ergonomic height for my particular desk and keyboard, I need a footrest so my feet aren't dangling. But that would be the case with any chair in my setup. The headrest is great, the arms are soft and comfortable, the back support piece is perfect, and everything can be adjusted as you sit in it. Rollers are quiet and stable on laminate floor.
The box was a good size and heavy enough (~32 lbs) to be awkward for an unfit middle aged lady to wrangle down apartment steps; however, the Amazon driver brought it straight to my door. The chair was carefully packed. Whoever wrote the assembly instructions (which are in English and Spanish) for this chair is some kind of technical writing genius -- I have never seen clearer or more easy-to-follow instructions. From slicing open the tape on the box to finished chair was 45 minutes for me. All you need for assembly is included -- also fabric gloves (for protecting your manicure? I guess?) and extra bolts and washers.
Super buy. If I'd known I could get such an affordable and massive upgrade from my previous garbage chair I would have done it long ago. Great value for the money. I rarely write reviews but all the reviews I read helped me make a great decision, so here ya go.
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Reviewed in the United States on October 29, 2024
★★★★★ 5
Comfort and ease
Color: Black, Style: With Headrest
Good chair all the functions work well I don’t really care for the armrest rotating but the fact they lift up an extend is nice. It’s comfortable. The lumbar is good. I don’t spend hours in a chair. It’s nothing fancy but definitely good for the price. Simple to put together seems durable enough.
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Reviewed in the United States on June 1, 2026