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Description
Mouse PECAM1 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 handling and requirements: Plasma: Collect the specimen using EDTA or heparin as an anticoagulant. Centrifuge at 1000g for 15 minutes at 2 8C within 30 minutes of collection. Remove the supernatant for testing or store at
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 handling and requirements: Plasma: Collect the specimen using EDTA or heparin as an anticoagulant. 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. 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 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 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 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 Platelet/Endothelial Cell Adhesion Molecule (PECAM1) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB converts to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of Platelet/Endothelial Cell Adhesion Molecule (PECAM1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Platelet/Endothelial Cell Adhesion Molecule ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Platelet endothelial cell adhesion molecule 1 (PECAM1) is a protein normally found on endothelial cells, platelets, macrophages and Kupffer cells, granulocytes, lymphocytes (T cells, B cells, and NK cells), megakaryocytes, and osteoclasts. The encoded protein is a member of the immunoglobulin superfamily and may be involved in leukocyte migration, angiogenesis, and integrin activation. PECAM1 on endothelial cells binds to the CD38 receptor on natural killer cells, allowing these cells to attach to the endothelium. It plays a key role in clearing senescent neutrophils from the body. | |||||||||||||||||||||||||||||||||
| 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 | plasma |
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4.7 ★★★★★
Based on 20 reviews
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Product Reviews
★★★★★ 5
Great product
Color: Black/Yellow Accent
Watch and service was great. I would recommend product to others.
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Reviewed in the United States on May 27, 2026
★★★★★ 4
No complaints after use
Color: Black/Yellow Accent
Always been a fan of Ironman watches especially the original. The negative display threw me off for a few days but getting used to it. Love the fit and size of it. Not to mention it's extremely light and you barely notice it after a bit. For under $40 if it malfunctions in a year or two I'll just buy another or one similar.
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Reviewed in the United States on November 4, 2025
★★★★★ 5
Bang for the buck
Color: Blackout
Let me previce this review by stating that I have had multiple fitness watches including Garmin suunto Timex and Casio. This watch has some limitations and I will get to them. The first thing is the design and functionality. The picture doesn't do this watch justice. It is sleek. Small enough to not be gaudy. But large enough to look appropriate regardless of your body size. I am 6' 180 lbs and its perfect. The functionality is great. Some watches require a college degree but this watch is simple to program and switch through modes. It has a lap counter countdown timer stop watch as well as time day date etc. It also has more features that don't interest me for fitness so I won't go into them. One complaint other reviewers have stated is with the black background it can be difficult to see display. To them I would reply the top right button is a light. Push it once to illuminate the display. Problem solved. Having a black background does make the display less visible. That's obvious to me but perhaps not everyone. With that being said I only have to use the light if I'm in a dark area. In normal room lighting it is easy to see. This watch does not have GPS capabilities which other watches I've had has. Add another zero to the price of this watch and you can have GPS too. I use this watch mainly for indoor fitness inc swimming and running on an indoor track and use my garmin outdoors. This is a great everyday watch and a great price. No issues no complaints.
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Reviewed in the United States on February 28, 2015
★★★★★ 5
6 years and still going
Color: Blackout, Color: Blackout
Owned for 6 years and still going strong. Worn daily, including showers. The only time I take it off is to sleep. The face is scratched up and some buttons worn smooth (I wear it on the inside of my wrist). A few tiny cracks are beginning to form on one of the wristband pieces. Replaced the battery after about 3 years. It has three alarms and I use them regularly. You can choose to set each alarm for daily, weekdays, or weekends. I also use the timer a lot. Indiglo gets used every morning and many evenings. You can disable modes that you never use, so you only see what you want. I never use chronograph, so I have that disabled. Incredible watch.
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Reviewed in the United States on May 11, 2026
★★★★★ 5
The watch of our era
Color: Blackout
Bought this ‘used’ but looks like only the box had some ‘damage’ but the watch was perfectly brand new. I’ve been wearing a timex Ironman watch since my tenth birthday. I’ve served in the Marine corps and wore one while on tour. I’ve worn them snorkeling and diving. I’m a mechanic and have worn it on my belt loop while working, and depending what I’m doing I’ll leave it on. I wear it in the shower, the pool, beach, kayaking, skydiving, mudding, surfing, fishing, shooting, and while relaxing in my recliner. I’ve only had one battery die, and it was easily replaced.
This is the watch of our era. Stop reading this review and buy one!
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Reviewed in the United States on April 27, 2026