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Description
Human PAR4 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 preparation and requirements: 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 and
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 preparation and requirements: 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 and mince the tissue. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1g of tissue sample to 9mL 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 collect the supernatant for analysis. Cell Lysis Buffer: Adherent cells should be gently washed with pre-chilled PBS, then trypsinized and harvested by centrifugation at 1000×g for 5 minutes. Suspension cells can be harvested directly by centrifugation. Collected cells should be washed three times with pre-chilled PBS and resuspended 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 freezing and thawing or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and collect the supernatant for analysis. 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 utilizes 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 Protease Activated Receptor 4 (PAR4). 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 Protease Activated Receptor 4 (PAR4) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Protease Activated Receptor 4 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Protease-activated receptor 4 (PAR4) is the fourth member of the PAR family and the third thrombin receptor identified and cloned in 1998. PAR4 is expressed in platelets and several other human tissues, such as the lung, pancreas, thyroid, testes, and small intestine, but is absent from the brain, kidney, spinal cord, and peripheral blood leukocytes. PAR4 comprises 385 amino acids and shares 27% and 30% sequence identity with PAR1 and PAR3, respectively. | |||||||||||||||||||||||||||||||||
| 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 | Tissue homogenates, cell lysates, and other biological fluids |
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4.3 ★★★★★
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Product Reviews
★★★★★ 5
My go to makeup removal wipes
Size: 1 Count (Pack of 25)
CeraVe makeup wipes are perfect for traveling. Plant based, non sticky, makes makeup removal a snap and best of all it first irritate my sensitive skin.
Open it up fully and simply wipe away your makeup. Place over eyes for about a minute to dissolve away mascara, eyeshadow and eyeliner. Truly it’s just that easy to easy to use. For me there’s no mess or fuss with using these wipes. They are a bit pricey, but you’re paying for a quality product.
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Reviewed in the United States on April 4, 2026
★★★★★ 5
Hydrating & Effective
Size: 1 Count (Pack of 25)
They are perfectly moist and leave my face feeling soft and hydrated without burning my skin like a lot of other makeup remover wipes do. They take off my makeup very easily without having to use much pressure.
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Reviewed in the United States on March 15, 2026
★★★★★ 5
Great at removing makeup
Size: 1 Count (Pack of 25)
I was really surprised at how well these wipes removed my full coverage makeup. I was looking for a “cleaner” option to replace the wipes I had been using for years. These were not dry, affordable, they were gentle on my skin and I only needed one wipe for my whole face. Will be buying them again.
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Reviewed in the United States on April 15, 2026
★★★★★ 4
MY go to for travelling
Size: 1 Count (Pack of 25)
CeraVe's Hydrating Facial Cleansing Makeup Remover Wipes are a popular choice in the skincare market, known for their dual functionality of removing makeup and cleansing the skin. Here's a review based on their performance and qualities:
Pros:
1. Gentle Cleansing: One of the standout features of these wipes is their gentleness on the skin. They are formulated with ceramides, which are essential for maintaining the skin's barrier and hydration. This means that even if you have sensitive or dry skin, these wipes are less likely to cause irritation compared to some other makeup removers.
2. Effective Makeup Removal: These wipes effectively remove makeup, including waterproof mascara and long-lasting lipstick. They are saturated enough to break down makeup without excessive rubbing or tugging, which is crucial for preventing damage to the skin.
3. Hydrating: As the name suggests, these wipes are hydrating, leaving your skin feeling moisturized and refreshed after use. This is especially beneficial if you have dry or dehydrated skin.
4. Non-Comedogenic: CeraVe products are generally known for being non-comedogenic, which means they are less likely to clog pores and cause breakouts. These wipes follow that trend and are suitable for most skin types.
5. Convenient for On-the-Go: The individual wipes are convenient for travel or for keeping in your purse or gym bag for quick touch-ups or post-workout cleansing.
Cons:
1. Scent: While these wipes are fragrance-free, I may still notice a faint chemical scent. It's not overpowering, but those with a sensitivity to scents might find it noticeable.
2. Environmental Concerns: Like many disposable makeup wipes, these are not the most eco-friendly option. If sustainability is a priority for you, you might consider alternative makeup removal methods, such as reusable microfiber cloths or cleansing balms.
In conclusion, CeraVe Hydrating Facial Cleansing Makeup Remover Wipes offer a gentle and effective solution for makeup removal and skincare. They are particularly well-suited for those with sensitive or dry skin due to their hydrating properties and ceramide-infused formula. While there are some minor drawbacks, such as the scent and potential packaging issues, they remain a popular choice among individuals seeking a convenient and reliable makeup remover that is also gentle on the skin.
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Reviewed in the United States on September 27, 2023
★★★★★ 5
Clean and care for your skin on-the-go
Size: 1 Count (Pack of 25)
Gentle face care with beneficial elements. I use these when I come in from outdoor activities, to wipe away sweat, pollen, dust. Leaves my skin conditioned, not dried out like others
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 6, 2026
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