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Description
Human PIEZO-2 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. Remove the supernatant for testing, or store at -20°C or -80°C, but avoid repeated freezing and thawing. Cell culture supernatant: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for testing, or store at -20°C or -80°C, but avoid repeated freezing and thawing. Pre-Test Preparation: 1. Remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Prepare a gradient standard working solution: Add 1 mL of universal diluent to the lyophilized standard. Let stand for 15 minutes to completely dissolve, then gently mix (concentration 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 500 μL of universal diluent to each tube. Pipette 500 μL of the 10 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 5 ng/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 Piezo-type mechanosensitive ion channel component 2 (PIEZO-2). After incubation and washing, the assay 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 Piezo-type mechanosensitive ion channel component 2 (PIEZO-2) in the sample. Absorbance (OD) is measured at 450 nm using a microplate reader to calculate sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Piezo-type mechanosensitive ion channel component 2 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | The mechanosensitive ion channel PIEZO-2 (PIEZO-2) is a protein encoded by the PIEZO2 gene. It has a homologous structure with three blades bent into a nanodome with a diameter of 28 nanometers. PIEZO2 is normally found in tissues that respond to physical touch, such as Merkel cells, and is thought to regulate light touch responses. | |||||||||||||||||||||||||||||||||
| 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 culture supernatant |
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4.2 ★★★★★
Based on 2016 reviews
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Product Reviews
★★★★★ 5
Great for bored dogs
Color: Flip Board, Size: Level 2
Great dog puzzle! My pup loves this, he did chew the peg portion once he got them out so they can be put back in. They are tough for small dogs to get out so I think he plotted it out.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on April 9, 2026
★★★★★ 5
Actually Holds up
Color: Knuckle Bone, Size: XX-Large
I must say I had low expectations when buying this bone based on prior experience of similar products. I'm happy to say I was pleasantly surprised. I bought this bone off Amazon in the beginning of December, I wanted to wait a while for my review to see if it would actually hold up. So far so good. There are several factors that set this bone above similar products.
First let me be clear that this is the Beef bone not the chicken etc. (I'm told there is a huge difference)
1. The first thing that impressed me is that my dog actually loves to chew on it. I know this may sound weird but I tried the puppy pack of nylabones and a bit larger one. He was interested in them but it didn't last and he wouldn't touch them anymore. This bone on the other hand is his absolute favorite. He carries it around everywhere. Even though it's beef flavored it doesn't have a noticeable smell or residue and it doesn't get slimly like a real bone.
2. This bone is extremely durable. I have a chew happy 10 month old lab puppy with razor teeth and majoring chewing skills. Trust me, my ruined shoes are proof :) It's been almost two months and I can just see where the bone has worn down. The down side to this is it is extremely heavy, not ideal to be leave around to step on, especially because of the way the bone breaks down. As they chew on it is almost peals away into tiny, tiny chunks. This is great compared to a kong stick that he had where he was swallowing large pieces of a rubber/plastic material. The downside is that as it "peals" it leaves the bone very sharp, with little spikes around where they chew. So far my dog doesn't seem to mind and it doesn't seem to make his mouth bleed or anything so that's a minor negative. Just don't stub your toe on it! Trust me!
Overall after trying multiple brand/types and flavors this is the only one that gets my seal of approval.
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Reviewed in the United States on January 29, 2012
★★★★★ 5
This is a great option for tough chewers
Color: Knuckle Bone, Size: XX-Large
This is a great option for tough chewers, although if your dogs are anything like ours, there is a procedure to ANY new toy no matter how "tough". If I gave them this Nylabone and they went unsupervised day one, they would wreck it and it'd be in the trash within the hour. I say this to tell you, if you have a top 10 bite strength aggressive chewer, that you shouldn't expect a miracle.
If instead, I give it, take it away when they are working it too hard or being particularly destructive, and then give it back- and do this a few times for a week or so, then they'll chew it in a less aggressive fashion- they'll enjoy it and it lasts a long time. I don't know what it is about the first few days, but when something is "new" it has to be destroyed.
When it's been in the house a few days, they are rough but not as bad. Like the ones listed "virtually indestructible" will last 6 months as long as we have multiple around. It will show damage, there will be shrapnel in the carpet that will scratch up your feet, but it'll be worthy of play still. I consider 6 months in our house worthy of 5 stars.
This has been true for 2 Dobermans and a German Shepherd- and all of them have proven ability to destroy these products day 1 if I don't behave appropriately on introducing the new toy. If your dog wrecks these in the first 10 minutes, I'd say you probably need to work on supervising new toy play, and taking it away when they become to aggressive. This is a training issue, it can be done!
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Reviewed in the United States on December 27, 2015
★★★★★ 5
Lasting longer than 6 months, longer than any bone to date.
Color: Mega Knot Bone, Size: X-Large
Finally a bone that has lasted longer than 6 moths so far. I know my dog is a chewer. Well worth the cost to not have to replace them as often.
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Reviewed in the United States on November 20, 2025
★★★★★ 5
Great quality
Color: Knuckle Bone, Size: XX-Large
Great quality. Our puppy is having a great time and it's holding up perfectly.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on November 9, 2025
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