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Description
Mouse uPA 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 anticoagulants and centrifuge them at 1000×g for 15 minutes at 2-8℃ within 30 minutes of collection. The supernatant can be tested or stored at -20℃ or -80℃, but repeated freezing and thawing should be avoided. 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 Plasminogen Activator and Urokinase (uPA). 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 Plasminogen Activator and Urokinase (uPA) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Plasminogen Activator, Urokinase ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Urokinase-type plasminogen activator (uPA), also known as urokinase, is a serine protease. Produced in the kidneys, it directly activates plasminogen to convert plasmin. It is the first generation of natural thrombolytic drugs extracted from urine. Urokinase protein was discovered by McFarlane and Pilling in 1947, but remains unnamed. Initially isolated from urine, it is also present in blood and the extracellular matrix of many tissues. The enzyme's primary physiological substrate is plasminogen, the inactive form (zymogen) of the serine protease plasmin. It directly acts on the endogenous fibrinolytic system, catalyzing the cleavage of plasminogen into plasmin. Plasmin degrades not only fibrin clots but also circulating fibrinogen, coagulation factors V and VIII, thereby exerting its thrombolytic effect. It also increases vascular ADPase activity, inhibiting ADP-induced platelet aggregation and preventing thrombosis. | |||||||||||||||||||||||||||||||||
| 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 | Serum, plasma, and other biological fluids |
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4.6 ★★★★★
Based on 27 reviews
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Product Reviews
★★★★★ 5
Lasts long
Color: Panda Bear - Large
Perfect for super chewer dogs.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 14, 2026
★★★★★ 4
Dachshund said “did I hear indestructible‽ Hold my ball”
Color: Grey Bear - Large
My 15lb Dachshund took this toy as a personal challenge.
He heard me tell his human brother that it’s indestructible on the inside and said “hold my ball” promptly proceeding to schmurder the textile aspect of this adorable toy.
I’d say it took him a total of 23 1/2 minutes to get to the center of this tootsie pop. I’m not even mad tho bc he had no interest in actually ingesting the outer part, and is totally into the inner toy, which is, for him, literally indestructible. It’s solid rubber, has a wild and fun bounce to it on multiple surfaces.
Zodiac rates the squeak 10/10. It’s great especially for when I’m on a call or talking to someone in real life and can’t immediately throw said toy or pay complete attention to him.
In conclusion:
Your dog will probably love it, ((suggest to just keep an eye on them during the schmurder process and make sure you grab all the pieces that come off so none are swallowed))
Depending on your tolerance for squeaker pitch and frequency of squeak, it's a 4/10 in annoyance mostly bc it’s not a long drawn out squeak ((I can not tolerate those toys, they get “lost on top on my fridge” until I get up the courage to throw them out without him seeing)), it’s quick and requires a bit of effort to produce the sound however it’s a very high pitch squeak so it’s all up to your preference. I personally try to buy toys I can tolerate my dog loving on bc when he gets a new toy, it’s his favorite thing for a long time, like, it sleeps on the bed with us for the first month.
Hope your fur child loves this as much as mine did.
Not sure I’ll do the whole two in one toy thing again only he got through it so fast.
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Reviewed in the United States on December 25, 2025
★★★★★ 5
LOVE BARK TOYS!!!
Color: Rad Herring - Medium
I got this toy for my 1yr old Husky/Heeler/(possibly)GSD (approx 42lbs) mix when she came to our home mid/late October of 2025 and the toy lasted until December 3rd 2025 when we had to assist her with opening the fabric to get the toy on the inside.
It was a great fun toy for her and the inside toys still is and she loves playing with it. The durability of both the outside and inside toys are great for super chewers! I was glad there was no stuffing because I had gotten her a plush toy once before and she destroyed that immediately. The fabric on this was durable enough to withstand her chewing yet soft enough that it didn’t hurt her teeth. The colors were vibrant and fun for both toys even though wear and tear kinda took some of the fabric color away. I would definitely buy this again once the inside toy is no longer safe for her, especially since the size was perfect for her too.
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Reviewed in the United States on January 12, 2026
★★★★★ 5
Good ball for strong chewers
Color: Cheeze Brawl - Large, Color: Cheeze Brawl - Large
My dog loves this ball except the outside fur gets gross really quickly. However, it’s easy to cutoff and underneath is a strong squeaky ball. It’s like two gifts in one. He’s a very heavy chewer and the ball lasts quite a bit.
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Reviewed in the United States on December 31, 2025
★★★★★ 5
Take the skin off and get the right size for your dog’s weight!
Color: Panda Bear - Large, Color: Panda Bear - Large
This review is specifically for the panda toy. My dog is a super chewer and will literally go through any normal toy, even the ones labeled for aggressive chewers, within a few hours at most. He’s about to be 10 years old and this is the first time he’s ever had a toy that lasted as long as it has. I wanna mention the outer layer he almost immediately chewed through. To me, it wasn’t worth the fear of him dealing through pieces so I just took the whole “skin” off. However, he’s safely had this for 2 months with no issues and heavy daily chewing until he finally broke it apart. So if you have a super chewer, make sure to get the size appropriate for your dog’s weight and take the “skin” off and you’re left with a quality toy that will stick around! Pictured is a set of 2 new toys of the same variety since he finally chewed through his last batch lol
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Reviewed in the United States on May 27, 2026