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Description
Mouse PSGL1 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: 1. 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: 1. 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. 2. 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. 3. 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. 4. 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. 5. 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. 6. 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 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 500uL of universal diluent to each. Pipette 500uL of the 10ng/mL standard working solution into the first EP tube and mix thoroughly to make a 5ng/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, biotinylated detection antibody, and HRP conjugate are sequentially added to microwells pre-coated with a P-Selectin Glycoprotein Ligand 1 (PSGL1) 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 P-Selectin Glycoprotein Ligand 1 (PSGL1) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse P-Selectin Glycoprotein Ligand 1 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | P-selectin ligand, also known as SELPLG or CD162 (cluster of differentiation 162), encodes PSGL-1, a high-affinity counterreceptor for P-selectin on myeloid cells and stimulated T lymphocytes. Therefore, it plays a key role in the ligation of these cells with activated platelets or P-selectin-expressing endothelial cells. The organization of the SELPLG gene is very similar to that of CD43 and platelet glycoprotein GpIb-α, with an intron located in the 5-prim noncoding region, a long second exon containing the entire coding region, and a TATA-less promoter. | |||||||||||||||||||||||||||||||||
| 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 | 0.15-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Serum, plasma, tissue homogenate, cell lysate, cell culture supernatant and other biological fluids |
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4.7 ★★★★★
Based on 588 reviews
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Product Reviews
★★★★★ 5
Highly recommended for any owner of a "Super Chewer."
Rating: ⭐⭐⭐⭐⭐ (5/5)
Title: Finally, toys that can actually withstand my Labrador’s jaws!
If you own a "power chewer," you know the struggle: you bring home a new toy, and within ten minutes, it’s shredded into a thousand pieces of foam and plastic. After going through countless toys that promised durability but failed within a day, I decided to give the WOWBALA 2-Pack Indestructible Dog Toys a try—and I am genuinely impressed.
The Durability Factor
The biggest selling point here is the material. These are made from high-density, food-grade nylon, and you can feel the quality the moment you take them out of the package. They are solid, heavy-duty, and have held up perfectly against my Lab’s relentless chewing. Even after a week of heavy use, there are only minor surface marks where his teeth have made contact, rather than the dangerous chunks that usually break off other toys.
Design & Engagement
The pack comes with two different shapes—a stick and a ring—which is a great variety. The curved, ergonomic design of the stick makes it very easy for my dog to hold between his paws while he chews, and the textured surface seems to really hit the spot for him. I’ve also noticed that it helps keep his breath fresh and his gums healthy since the texture acts almost like a toothbrush while he plays.
The "Milky" Scent
One thing I was initially skeptical about was the "milky scent." I wondered if it would be overpowering, but it’s actually quite subtle. It was strong enough to immediately pique my dog’s curiosity right out of the box, but it doesn't leave a weird smell on my carpet or hands. It’s clearly effective, as he keeps coming back to these toys even when he has others scattered around the room.
Peace of Mind
Beyond just being tough, I appreciate that they are non-toxic and dishwasher safe. After a long day of outdoor play in the mud, being able to just toss these into the dishwasher and have them come out clean and sanitized is a huge convenience.
Final Verdict
If you have a medium to large breed dog that loves to destroy everything in sight, these toys are well worth the $15.99 investment. It’s rare to find a product that lives up to the "indestructible" label, but these truly do. They’ve been a lifesaver for keeping my dog occupied while I’m working from home, and it’s a relief not having to play "clean up the debris" every evening.
Pros:
Extremely durable high-density nylon.
Great value (2-pack).
Easy to clean (dishwasher safe).
Engaging design for teething, training, and boredom relief.
Safe and non-toxic.
Cons:
Because they are hard nylon, be careful using them indoors on hardwood floors—they can be a bit noisy when dropped!
Highly recommended for any owner of a "Super Chewer."
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Reviewed in the United States on May 19, 2026
★★★★★ 5
These are way better than I expected and hold up to our aggressive chewer spectacularly!
I got these specifically for our newest pup (an 8 month old lab/pitbull mix--she's a big puppy, lol) who is obsessed with toys and chewing. She has a tendency to go through her toys quick, both chew toys and soft ones, so I wanted something that would last a bit longer for her because I can tell she gets a bit sad when we have to trash one that's hit its lifespan. So far, these have been a big hit! They're the perfect size for her to pick up, the chicken claw-style texture is great for both keeping her interested and giving her some grabbing options, and they do seem to hold up to their claim of being made for aggressive chewers! There's obviously some wear, but it looks like expected chew toy wear; no claws have snapped off, and her chew marks look relatively minimal compared to what she's done to other chews. I also appreciate that these came in a two pack; we have a smaller dog, and while these are technically big for him, he enjoys having one to play with and chew on as well. These have been a pleasant surprise and I actually think I'll come back to buy them again whenever they're spent! Definitely a worthy purchase!
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Reviewed in the United States on June 2, 2026
★★★★★ 5
Creepy chicken feet, made for big dogs and Frenchie approved
These chew toys have been a hit with our dogs.
We have three French Bulldogs who firmly believe they are ferocious wolf-sized predators, and these toys have held up surprisingly well to their daily chewing sessions.
The chicken foot shape keeps them interested, and the durable nylon construction has stood up to much more abuse than many other toys we've tried.
The toys are easy for the dogs to grip and carry around, and I appreciate that they're simple to clean when they inevitably get covered in slobber. They've also helped keep the dogs occupied and redirected some of that endless Frenchie energy away from furniture and other household items.
Overall, these have proven to be durable, entertaining chew toys that are Frenchie-approved.
Although, based on the dramatic growling and victorious prancing around the house, you'd think they personally hunted and defeated a giant prehistoric chicken
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Reviewed in the United States on June 14, 2026
★★★★★ 1
Waste of $ smell of bacon went away in 1 day
Dont waste your $ smell of bacon/meat goes away after 1 day. Waste of money & the groove to put peanut butter DOESN'T HOLD ANY PEANUT BUTTER !
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Reviewed in the United States on June 4, 2026
★★★★★ 4
Creepy Little Chicken Feet… But My Dog Is Obsessed
Creepy Little Chicken Feet… But My Dog Is Obsessed
I am not even going to lie, when I first opened these WOWBALA dog toys, I had to pause for a second because why do they look like actual little chicken feet? 😂 They are definitely a little creepy at first glance, but apparently my dog does not share my human judgment because she went absolutely crazy over them.
These come as a 2-pack, and they have a bacon scent that is very much not for me, but clearly made perfect sense to her. The second she got one, it became her new prized possession. She carried it around, chewed on it nonstop, and I am pretty sure she even tried to hide it somewhere in the backyard like she had discovered buried treasure.
What impressed me most is how well these hold up. My dog is not gentle with toys at all. She chews like she has a full-time demolition job, and somehow these still look basically new from a normal glance. Up close, yes, you can see where she has been working on it, but it has not broken apart, shredded, or left those annoying little toy pieces all over the floor. If you have ever bought a chew toy that turns into confetti within 20 minutes, you already know why that matters.
The nylon feels solid and heavy-duty, and I like that it is made with safe, non-toxic material since this is something she keeps in her mouth for long periods of time. It also keeps her busy, which is a huge win because bored dogs will find their own entertainment, and usually it is something expensive or inconvenient.
So yes, the chicken feet design is a little weird. Yes, the bacon smell is definitely for the dog and not the humans. But my dog is obsessed, it keeps her entertained, and it survives her chewing like a champ. That makes this a solid win in my house.
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Reviewed in the United States on June 2, 2026