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Description
Human CCDC134 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 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 capture antibody against Coiled-Coil Domain Containing 134 (CCDC134). After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase and to yellow by acid. The intensity of the color is positively correlated with the amount of Coiled-Coil Domain Containing 134 (CCDC134) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Coiled-Coil Domain Containing 134 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Frizzled-coil domain-containing protein 134 (CCDC134) is a well-characterized secreted protein that acts as an immune cytokine and exhibits potent anti-tumor effects by enhancing CD8+ T cell-mediated immunity. Diseases associated with CCDC134 include osteogenesis imperfecta type Xxii and osteogenesis imperfecta type Xvii. | |||||||||||||||||||||||||||||||||
| 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, cell lysates and other biological fluids |
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★★★★★ 5
Silky and Soothing
Color: Cream, Size: 2 Pack Queen（20"X 30"）
I thought I was getting 1 of the queen sized but I got 2. I got these for my Pomeranian's fuzzy pillow. He's always hot and these are very cooling and very good quality. He loves them and are very comfortable for him. They totally transformed his shaggy old pillow. I used the larger body pillow covers to replace the covers I had. They matched my bedspread perfectly. I use 2 body pillows along the exterior wall in my bedroom where there is cold coming through the floor in the winter. I'm sure I will enjoy them in the summer when it's hot too!
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Reviewed in the United States on January 10, 2026
★★★★★ 5
Comfortable and stays cool
Color: White-cooling Basic, Size: Queen(Pack of 1)
Great pillow. When i opened it, i noticed a strange smell, i assumed its the smell of whatever they use to make the pillow stay cool. You have to fluff it for a good amount of time to make sure the cooling agent spreads well in the foam. Is what i assumed atleast. Its a perfect amount of softness for me. Pair it with some bamboo pillow cases and you got a good night of sleep
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Reviewed in the United States on May 9, 2026
★★★★★ 5
BEST, MOST COMFORTABLE PILLOWS EVER!
Color: White-cooling Basic, Size: Queen(Pack of 2)
I've used others.BEST PILLOWS EVER! Firm, cooling and supportive. LOVE THESE!
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Reviewed in the United States on May 28, 2026
★★★★★ 4
Comfortable and Supportive, but Adjust the Filling
Color: White-cooling Basic, Size: Queen(Pack of 2), Color: White-cooling Basic, Size: Queen(Pack of 2)
I’m very happy with this pillow. The size is big and it feels soft while still providing good support, perfect for midnight scrolling on TikTok.
One thing to keep in mind is that it can feel a little too tall if you use all of the filling that comes with it. The good news is that you can easily adjust the amount of filling to find the height that works best for your sleeping position. Once I customized it, it was much more comfortable.
Overall, great quality, comfortable, and worth the purchase.
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Reviewed in the United States on June 3, 2026
★★★★★ 5
BEST PILLOWS EVER!!!
Color: White-cooling Basic, Size: Queen(Pack of 2)
I am very picky with my pillows because I have a bad neck and I can never seem to find a good pillow. But I tried this pillow because it had everything I was looking for and the price was great! You get two of them and they are absolutely awesome.! normally I would need to lay on two pillows to try and get comfortable but with these pillows I just lay on one and I wake up with no neck pain. They really have a nice cooling effect. They actually puff up immediately after opening them. All you have to do is just shake them up and the pillow just puffs straight up.. It comes with extra cushioning if you want it to make it firmer and if you don’t want it as firm as it is, you can actually take some out. It really is such a beautiful pillow. It’s perfect in every way to me being such a picky pillow person.
You just lay your head down on it and it forms around your neck and your head and when you get up, it goes right back to its normal form. It doesn’t get flat and I love it. I completely recommend this to anyone who is a person who has a hard time finding a really good pillow.
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Reviewed in the United States on April 12, 2026