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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 22 reviews
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Product Reviews
★★★★★ 4
Stunning
Size: 1.7 Ounce (Pack of 1)
Good lord this is a beautiful mineral sunscreen. And at a reasonable price. It has a mouse-like consistency, is light and airy dries down to a satin-like finish after 15 or so minutes. It has a primer like feel to it which some might not like but this is perfect for those of us with combo/oily skin.
I’m fair/light skinned and the color is a perfect match. Gives me a light no-makeup makeup look. Works well under make-up as long as you let it set first.
One thing that's a con and has me on the fence about daily use is that because it's so thick and silicone like, you can't put the full 1/4 tsp on your face right away as it's just too much. For the 2 finger application I have to put it on in stages, and let each finger length dry down before adding the next, otherwise it's too much product to do all at once. Takes longer in the morning which isn't ideal but given everything else it has going for it it's still workable for me. I'll see over time if I want to repurchase.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 7, 2025
★★★★★ 5
Light and Feels like whipped butter - Tinted
Size: 1.7 Ounce (Pack of 1)
Love the tinted type of this. Soft and creamy to put on my face and feels lighweight. Color is not overwhelming.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 1, 2026
★★★★★ 5
Excellent sunscreen, my favorite
Size: 1.7 Ounce (Pack of 1), Size: 1.7 Ounce (Pack of 1)
In search for a perfect tinted sunscreen I have tried just about every brand of tinted mineral sunscreen that’s available here on Amazon, so you might recognize me from other reviews. I have a sensitive skin and can’t tolerate chemical sunscreen. I also have red and bumpy skin on my cheeks due to rosacea, so I need a tint to cover redness. I’m really glad I finally decided to give Sunbum tinted sunscreen a try because it quickly became one of my favorites. My other favorite is MD Solar Science BB cream in shade Light. These two are quite similar, both are very silicone like, due to a great amount of dimethicone. They feel like primers and because of that are very sticky and hard to wash off your fingers. But both look very natural and skin like. MD Solar Science (light shade) is slightly darker while Sunbum is pretty pale but not in an unhealthy way. My skin is a bit darker than Sunbum shade right now. Yet it covers my rosacea and evens out my skin tone just enough. It’s not drying but also not shiny, and because of that it doesn’t magnify wrinkles, which is a big plus: I’m tired of tinted sunscreens highlighting the wrinkles I have that are not even noticeable without them.
Sunbum is slightly lighter and provides a bit less coverage than MD Solar. It also doesn’t have much zinc, which was a point of criticism from popular bloggers. Because of that it might not be perfect for everyday wear. But if you want to have the most natural “no make up” look and you’re light skinned, this is a great choice on the days when you’re not expecting to be under too much sun. I use retinol on my forehead at night and I didn’t get sunburnt when using this sunscreen. So the only concern here would be the gradually accumulated damage from the long UV-A rays.
Another thing I like is that unlike MD Solar, Sunbum doesn’t contain vitamin C and therefore doesn’t irritate my sensitive skin. It’s also less expensive. I will keep repurchasing.
Note: in the unfiltered photo attached here, you can see the sunscreen after a 5 hour wear, that included eating, touching my face etc, so it’s a bit worn off and the redness on my cheeks is more visible. Even when freshly applied it didn’t cover redness completely because of being light coverage sunscreen.
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Reviewed in the United States on July 20, 2023
★★★★★ 3
Decent but not the best
Size: 1.7 Ounce (Pack of 1)
I like that this sunscreen is matte and tinted, but that’s about it. I have pretty oily skin and the mattifying effect only lasts for about 6-7 hours.
Unfortunately this product doesn’t absorb very well either. It tends to accumulate in any wrinkles and also tends to sit on top of any sort of facial hair or areas of dry skin.
A much better product, that absorbs very well and controls oil literally all day long, is Trader Joe’s mattifying facial sunscreen in the sea foam green tube. It isn’t tinted like Sun Bum but it beats it in every other category, especially in price.
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Reviewed in the United States on April 29, 2026
★★★★★ 5
Holy grail face SPF - mineral, tinted and matte
Size: 1.7 Ounce (Pack of 1)
Title says it all. This is my holy grail face SPF. It’s mineral and non-toxic. It’s lightly tinted so it looks beautiful on your skin alone and helps even out your skin. It’s also matte so it wears well under makeup. It’s moisturizing and applies and blends smoothly onto the skin. Amazing product. I have it on subscribe and save.
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Reviewed in the United States on September 10, 2025
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