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Description
Mouse GPC5 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: Serum, plasma, 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
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: Serum, plasma, 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 freeze-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 freeze-thaw cycles 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 1000×g for 20 minutes, and collect the supernatant for analysis. Preparation for the Assay: 1. Remove the reagent 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 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 glypican 5 (GPC5) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase and to yellow by acid. The intensity of the color is positively correlated with the amount of glypican 5 (GPC5) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse glypican 5 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Glypican-5 (GPC5) is a protein encoded by the GPC5 gene. Heparan sulfate proteoglycans on the cell surface are composed of a membrane-associated protein core punctated by a variable number of heparan sulfate chains. Members of the glycoprotein-related integral membrane proteoglycan family (GRIPS) consist of a core protein anchored to the cytoplasmic membrane via glycosylphosphatidylinositol linkages. These proteins may play a role in controlling cell division and growth regulation. | |||||||||||||||||||||||||||||||||
| 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, tissue homogenate, cell lysate and other biological fluids |
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4.7 ★★★★★
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Product Reviews
★★★★★ 4
Nice
Size: 3T-4T, Unit Count: 112
Good pull ups, it could get a little linty but it fits perfectly on my toddler. It stretches which I love. I do love how I could strap and unstrap the sides of the pull up. The designs on the pull up are cute. It holds a lot of pee and soaks it up very well. It’s comfortable on my toddler and the pee and poop doesn’t leak out of it. It is worth the money.
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Reviewed in the United States on November 18, 2025
★★★★★ 5
The Fast & the Curious- Potty Drift
Size: 5T-6T, Unit Count: 60
Who knew toddler potty training required both a PhD in fluid dynamics and Jedi mind tricks? These Pull-Ups prove the science part: leak-proof, absorbent, and surprisingly soft. The toddler-psychology part? That’s a battle of wills.
Our little one was a die-hard fan of Huggies diapers—too loyal, in fact. We begged, we bribed, we pleaded: “Please, let’s move on!” Resistance was fierce. So we went stealth mode, sneaking Pull-Ups on at night like parents on a covert mission.
Credit to mom—she cracked the code and finally got the kid interested in the toilet. Suddenly, the “potty rush” moments became less of a flood and more of a sprint. Diapers were too slow to peel off; Pull-Ups gave our little sprinter the speed boost they needed.
They’re soft, rash-free, and practically leak-proof (unlike that other brand we don’t speak of). Pro tip: size up for easier toddler self-removal.
The result? A drier, happier kid, less laundry for us, and the toilet finally earning MVP status. If you’re ready for your toddler to graduate from Diaper University, this is the diploma. Bonus: you’ll save money too.
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Reviewed in the United States on September 3, 2025
★★★★★ 5
Mom of 3 recommend!
Size: 5T-6T, Unit Count: 14
I'm using these again for the 3rd time and they're great as always. My 3rd is still having some issues at night time with bed wetting, and as they get older the bigger the mess, so these truly hold up to a full bladder. And they don't smell by morning like some diaper brands. I'm usually against pull ups, but these are great for up and down ease of use and that added protection when it comes to potty training. I love the bigger size inclusion for larger children which we struggle with for comfort compared to other brands.
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Reviewed in the United States on February 14, 2026
★★★★★ 5
Potent and Pure — You Can Feel the Difference
Flavor Name: Extra Strong, Size: 3.4 Fl Oz
This black seed oil is definitely strong — you can tell it’s pure and freshly cold-pressed from the first taste. The flavor is bold but clean, exactly what you’d expect from a high-quality oil. I appreciate that it’s ethically sourced and bottled in glass instead of plastic, which helps preserve its freshness.
I’ve noticed a difference in overall energy and digestion after adding it to my daily routine. It’s not the easiest oil to take straight, but it’s worth it for the results. Solid value for the quality and strength.
Black seed oil has become one of my go-to oils for skincare. It absorbs quickly without leaving a greasy film and helps calm irritation, dryness, and redness almost instantly. I like to mix a few drops with my regular moisturizer or apply it directly at night — it leaves my skin soft, hydrated, and with a healthy glow by morning. The natural antioxidants and fatty acids make it great for restoring balance, especially if you deal with uneven tone or occasional breakouts. It’s a strong oil, so a little goes a long way, but the results are worth it — smoother, clearer, and healthier-looking skin.
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Reviewed in the United States on October 12, 2025
★★★★★ 5
Said to have healing qualities - high quality and pure in nice package - but could use a drip spout
Flavor Name: Extra Strong, Size: 3.4 Fl Oz
Super high quality - pure - blackseed oil. If I were to change anything, I would want it in a container that I could control the flow out of. That said - I use this on my knee (which is giving me some trouble these days) - hearing from a co-worker that in another country, this oil used with some prayers, is known for healing. I can't vouch for that - but I do use it as a matter of hoping the oil might get into my joints and help my knee hurt a little less some days. The packaging is great, and I love the bottle. I would just encourage them to put a 'drip spout' on the bottle - because if you know anything about oil, it finds it's way down the sides of the bottle if you aren't careful. Easy clean up since it's a natural substance though. It does have an interesting 'gun oil' smell - to me. It's not bad in any way. And it also seems to absorb quickly into my skin. It is on the brown-ish side, and if you rub your skin against material before it dries - well just remember - it's 'oil'...and may discolor what you touch.
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Reviewed in the United States on May 16, 2025