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Description
Human PCII 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 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. 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. 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. 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. 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 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 7 EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 20 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 10 ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves directly as a blank well; there is no need to aspirate the liquid from the penultimate tube. See the figure below for details. 3. Preparation of Biotinylated Antibody Working Solution: 15 minutes before use, centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration using universal diluent (e.g., 10µL concentrate + 990µL universal diluent). Prepare immediately before use. 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 enzyme conjugate are sequentially added to microwells pre-coated with a Procollagen II (PCII) capture antibody. 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 II (PCII) 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 II ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Procollagen is the precursor of tropocollagen and is the form secreted into the extracellular space. Procollagen is composed of three pre-α-chain helices, each with an N-terminal propeptide of 150 amino acid residues and a C-terminal propeptide of 250 amino acid residues. PC II is a type of procollagen. | |||||||||||||||||||||||||||||||||
| 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 | 0.31-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.0 ★★★★★
Based on 19 reviews
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Product Reviews
★★★★★ 5
Great Value for a ball my dog loves!
Color: S6-muti-color
I wish these ball were in the subscription program! My dog gets sooo excited by the new ball, but her interest in things lasts for a few days. She kills the squeak within 11minutes, but the squeaker stays in place rather than becoming a chocking or swallow hazard, and she still loves to chew it. Next, we must throw the ball approximately 723 times the first day, possibly 496 times on day 2, and just 3- 4 on day 3, at which point she will chase the ball before hollering "it's over here if you need it," while she checks on the chipmunk den. We currently have 17 of these balls in our yard (because we have given several leftovers to a less discriminating doodle next door). These balls hold up really well, get her extremely active for the first couple of days, and are a much cheaper than doggie daycare (after an active morning with a new ball, she is happy to chew on it and sleep much of the day). I always have these on hand!
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Reviewed in the United States on November 22, 2025
★★★★★ 4
Pieces chewed off
Color: white & green
My Aussiedoodle loves it. I throw it down the hallway, it bounces crazy, and he chases it. The only complaint is that he can't chew much of the ball part. So now I have to have it put away and brought out for special playtime.
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Reviewed in the United States on April 9, 2026
★★★★★ 5
Hyper Pet Tennis Balls for Dogs: Because Every Dog Deserves to Play Fetch Like a Pro
Color: Green, Size: Regular 2.5" - 12 Pack
If your dog had a bucket list, fetching tennis balls would probably be near the top. And if that’s the case, then the Hyper Pet Tennis Balls are their VIP ticket to Fetch World. These little balls of joy are the real deal, perfect for small breeds that are living their best life chasing after something that probably doesn’t need to be chased but is just so throwable.
The 12-pack means you can never lose a ball again...unless, of course, it gets lost in a mysterious “dog dimension” where all the socks go. These 2.5-inch tennis balls are the right size for those dogs that aren’t quite large enough to play with the big leagues. Think of them as the perfectly sized appetizer for the full-on Fetch Feast. Plus, they’re made from non-toxic material, so your pup can gnaw on them all day without worrying about a tummy ache (unless they’re overzealous and gobble up the whole ball...don’t worry, I won’t judge).
If your dog doesn’t love fetch yet, they will after these balls come into their life. They’re designed to be durable enough for constant chomping while still offering that classic “bounce, roll, and fetch” action that gets dogs going.
Pros:
• 12 balls = endless fetch sessions (until the dog gets tired, but let’s be honest, that’s probably never).
• Perfect size for small breeds—no choking hazards, just good ol’ fun.
• Non-toxic materials for guilt-free playtime.
• They squeak. And we all know that squeak is like doggy kryptonite.
• Durability means no more fetching balls that break after three tosses (we’ve all been there).
Cons:
• They might not hold up as well with very heavy chewers (but don’t worry, your dog won’t know the difference until they’ve already swallowed half of one).
• They don’t come in tennis-ball-flavor. If they did, I think we’d all be out of a job.
If you’re looking to upgrade your dog’s fetch game (or just keep them entertained while you catch up on your own Netflix binging), these Hyper Pet Tennis Balls are a solid choice. Plus, your dog will be too busy fetching to judge your questionable snack choices.
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Reviewed in the United States on December 13, 2024
★★★★★ 5
Prefect for smaller dogs to play
Color: Green, Size: MINI 1.75" - 12 Pack
The only balls my sato mix will fetch! They are a bit smaller than regular balls and are more comfortable in my dogs smaller mouth (she is about 20 lbs). Great bounce and durability, highly recommend!
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Reviewed in the United States on May 19, 2026
★★★★★ 5
Perfect Size - No Squeakers (yay)
Color: Pink, Size: MINI 1.75" - 4 Pack
My 9 lb. Chi-Mix adores her little toy balls. Perfect for her small size (or a medium dog) and full of bounce. I didn't want squeakers because I knew it would drive me nuts. No squeakers in these balls. Perfect training tool to teach fetch. Perfect toys to have play time with your pooch.
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Reviewed in the United States on April 14, 2026