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Description
Human Cyclin D1 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 freeze-thawed. Finally, centrifuge the homogenate at 5000×g for 5-10 minutes and remove the supernatant for analysis. Cell culture supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for analysis or store at -20°C or -80°C, but avoid repeated freeze-thaw cycles. Pre-Assay Preparation: 1. Remove the reagent 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 stand for 15 minutes to completely dissolve, then gently mix (concentration 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 seven EP tubes and add 500uL of universal diluent to each tube. Pipette 500uL of the 20ng/mL standard working solution into the first EP tube and mix thoroughly to make a 10ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube is used 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 cyclin D1 capture antibody. 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 cyclin D1 in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Cyclin D1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Cyclin D1 is a protein encoded by the CCND1 gene. The CCND1 gene encodes the cyclin D1 protein. This gene is located on the long arm of chromosome 11 (band 11q13). It is 13,388 base pairs long and translates to 295 amino acids. Cyclin D1 is expressed in all adult tissues, except cells derived from bone marrow stem cells (both lymphoid and myeloid lineages). Its overexpression has been shown to be associated with early cancer onset and tumor progression, and it can contribute to tumorigenesis by increasing anchorage-dependent growth and angiogenesis through VEGF production. | |||||||||||||||||||||||||||||||||
| 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 | 0.312-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenate, cell culture supernatant |
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4.1 ★★★★★
Based on 814 reviews
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Product Reviews
★★★★★ 5
It works! Quality pillow.
Size: Standard (1 Count), Color: Original White (Cooling)
This does actually provide a cooling effect, and it's very comfortable! The accompanying literature is helpful too. I believe this to be a quality pillow for the price.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 2, 2026
★★★★★ 5
Most comfortable pillow I’ve ever slept on
Size: Queen (1 Count), Color: Crescent White(cooling)
This is the most comfortable pillow I have ever slept on. It doesn't mash down like other pillows. Cradles my neck perfectly plus it’s cool. I broke my neck a couple of years ago and this pillow feels very good and comfortable. Definitely recommend to everyone. Ive used it since May and it’s retained its shape.
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Reviewed in the United States on June 6, 2026
★★★★★ 4
Smell
Size: Queen (2 Count), Color: Original White (Cooling)
The fact that there are not more comments on the smell of these pillows has me worried about some of yall. Listen, I LOVE the weight, and the firmness, it’s perfect for that. However, these pillows are RANK. They have a very strong mildew-ish smell that lingers. I took the outer cover off and washed it and let the pillow air out for a few days, the smell is still very strong when you press your face against it, which one does because it is in fact designed for that purpose. It smells so bad. It’s nauseating. I can smell it through two pillowcases and blanket that I threw on top of them hoping an extra layer would trap the smell. I’ll bake them in the Texas sun tomorrow and hope that helps, because otherwise these are the perfect pillows for someone who wants to feel like they are hugging a torso but can’t stand the sound/feel of someone else breathing. I don’t know…4 stars, if the smell goes away I’ll come back.
Update: many many days later. Either they don’t really smell anymore or I got use to it. They’ve held their firmness and shape well. I like them.
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Reviewed in the United States on March 9, 2026
★★★★★ 5
Actually Cooling Pillow
Size: Queen (1 Count), Color: Original White (Cooling)
This is a really nice pillow, I was worried it would be too soft or too firm but it's just right and it's actually cooling, it feels cool every time I touch it. I've noticed an improvement in sleep since using it. Would recommend!
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Reviewed in the United States on May 27, 2026
★★★★★ 5
Repurchased in King Size – Still Love These Cooling Pillows
Size: King (2 Count), Color: Original White (Cooling), Size: King (2 Count), Color: Original White (Cooling)
After using the queen-size version of these pillows for over a year, we purchased the king-size set when we upgraded to a king mattress. Both my husband and I really like them.
The cooling feature works well, and we like that each side feels different—when I run cold, I use the softer bamboo side, while my husband prefers the cooler side since he tends to sleep hot. They fit well in our king-size pillowcases and feel supportive in all sleeping positions, whether I’m on my stomach, side, or back.
I also appreciate that you can remove some of the filling to adjust the firmness. The covers are washable, which makes them easy to maintain. Overall, we’re very happy with this repurchase and the long-lasting comfort.
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Reviewed in the United States on April 15, 2026
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