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Description
Human PINP 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 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 procollagen I N-terminal propeptide (PINP). 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 procollagen I N-terminal propeptide (PINP) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Procollagen I N-Terminal Propeptide ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Peptides are cleaved from procollagen by proteases to form collagen. The concentrations of these two peptides in the bloodstream reflect the rate of type I collagen synthesis. Procollagen type 1 N-terminal peptide (PINP) is considered the most sensitive biomarker of bone formation. PINP exhibits diurnal variations, with higher values at night. PINP is metabolized in the liver. | |||||||||||||||||||||||||||||||||
| 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 | 0.15-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.1 ★★★★★
Based on 28 reviews
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Product Reviews
★★★★★ 5
Feels Like Sleeping on a Cloud — One of My Favorite Bedsure Products Yet
Color: Black, Size: King
I’ve had really good experiences with Bedsure products over time, so whenever I get the chance to try another one I usually do, and this mattress topper definitely did not disappoint. This one went on my mom’s king size bed and the difference was noticeable immediately.
The topper has a soft, cloud-like feel while still giving enough support underneath so you don’t just sink straight through it. It added a much more comfortable layer to the mattress and made the bed feel cozy and inviting without being overly heavy or hot.
I also appreciated how easy it was to put on. The deep pocket design fit securely over the king mattress without struggling to stretch the corners, and once it was on, it stayed in place well through the night.
The material feels breathable and smooth, which is especially nice for warmer nights. My mom commented that she slept really well with it and woke up feeling more comfortable than usual, which honestly says a lot.
Overall this feels like a high-quality mattress topper that makes a bed instantly more comfortable. Bedsure continues to be one of those brands I trust for soft bedding and reliable quality.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 23, 2026
★★★★★ 2
Not Worth the Price
Color: Black, Size: Queen
I expected more. The images are a bit misleading with the amount of fluff and thickness the product actually has. Despite following the instructions, it’s pretty flat.
It had a funky chemically smell to it, so I sprayed with some Febreeze.
Honestly in my opinion I don’t think it’s worth $60. I think if you want something similar, go to Marshals or TJ Maxx.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 30, 2026
★★★★★ 5
Makes my camp mattress so much better.
Color: Black, Size: Full
I'm in a small motorhome and this has helped my sleep quality a ton. I got rid of my big mattress because the 2 camp mattresses I have are smaller in size, but a bit dense than the mattress I had. Uncomfortable for anyone that sleeps on them for long periods and when I come from biking or hiking the pain was making it hard to sleep. These have improved my sleep quality immensely. It also, to my surprise fit well, the cushions are only a few inches. These are up to 18, but the edge is nearly a perfect fit at the start. The straps keep it from moving. The length and width using 2 of the bed rolls is nearly perfect. On mattress the deeper part of the pocket, would fit perfectly.
The quality feels great, lightweight and washable, this is one of those items I wouldn't have likely ordered, but trying it is near priceless for my usage. Couldn't imagine how good it must be on a bed.
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Reviewed in the United States on May 27, 2026
★★★★★ 5
Ultra-Light, Cooling & Comfortable – Great for Night Sweats!
Team Name: Cooling Comforter, Size: Full/Queen(90"x90"), Color: 01dark Grey
The DOWNCOOL Cooling Comforter Set (Queen Size 90”×90”) is a fantastic choice if you tend to sleep hot. The comforter uses advanced cooling fabric technology that absorbs body heat and helps reduce skin temperature by a few degrees, making it ideal for summer or warm climates.
The material feels cool to the touch, lightweight, and breathable, so you stay comfortable without feeling heavy or sweaty. It also features moisture-wicking and quick-dry properties, which help keep the bed fresh throughout the night.
The set includes 3 pieces (comforter + 2 cooling envelope pillowcases), and the pillowcases add an extra cooling effect around your head and neck. The fabric is soft and skin-friendly (often OEKO-TEX certified), making it safe for everyday use.
It’s also machine washable and durable, maintaining its cooling performance even after multiple washes, which makes it practical for long-term use.
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Reviewed in the United States on March 18, 2026
★★★★★ 5
Cool and comfy
Team Name: Cooling Comforter, Size: King(104"x90"), Color: 01grey
So comfy, lightweight, soft and yes, cool to the touch. Doesn’t stay cool though for long exposure but great for sleeping with animals or others.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on January 2, 2026
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