SKU: 71446778713

Human uPAR ELISA Kit

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Description

Human uPAR 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. 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.

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, Urokinase Receptor (uPAR). 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, Urokinase Receptor (uPAR) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration.
Source Human
Synonym Human Plasminogen Activator, Urokinase Receptor  ELISA Kit
Detection Type Double antibody sandwich method
Composition
Name 9 6 T  match   set remark
Pre-coating 96 Well plate 8 Hole ×12 Strip without
Standard 2 branch
Dilute as per instructions
Universal diluent
2×20mL
without
Concentrated biotinylated antibody ( 100× )  
120uL
Dilute as per instructions
Concentrated enzyme conjugate ( 100× )
120uL
Dilute as per instructions
20× Washing liquid
2×10mL
Dilute as per instructions
Bottom thing ( TMB )
10mL
without
Stop liquid
6mL
without
Sealing film
4 Zhang
without
Instructions
1 Share
without
Background The urokinase-type plasminogen activator receptor (uPAR), also known as the urokinase receptor or CD87 (Cluster of Differentiation 87), is a protein encoded by the PLAUR gene in humans. It is a multidomain glycoprotein tethered to the cell membrane by a glycosylphosphatidylinositol (GPI) anchor. uPAR was originally identified as a saturable binding site for urokinase (also known as uPA) on the cell surface. uPAR is composed of three tandem LU domains, a member of the three-finger protein family. The structure of uPAR in complex with a peptide antagonist and its native ligand, urokinase, has been solved by X-ray crystallography. uPAR also interacts with several other proteins, including vascular proteins, uPAR-associated protein (uPARAP), and the integrin family of membrane proteins.
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.
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, tissue homogenates and other biological fluids
Shipping Notes
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Exchange/Return Notes
  • We offer a 30-day return/exchange service after receiving.
  • Final sale items are not eligible for returns or exchanges.
  • To process your return/exchange, please contact us at [email protected]
  • Please click here for more details>>> Return & Exchange Policy
SKU: 71446778713

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4.6 ★★★★★
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Mo
Dallas, US
★★★★★ 5
A Hardy Toy for Chewers
Color: Bule Octopus, Size: For Larege Dogs, Color: Bule Octopus, Size: For Larege Dogs
I must say I was very surprised with the durability of this toy. We've had it for 3 days and not one puncture hole. My rescue is a destroy chewer that doesn't care for Hardy chew toys unless it squeaks. While the squeaker isn't the best, he still loves it. We put peanut butter in the grooves for him and he goes to town. Will purchase again should he ever destroys it, worth the price!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 8, 2026
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Verified Purchase
Jeffrey Brown
Fort Morgan, US
★★★★★ 5
Durable toy for a great price! Will buy again.
Color: A Red, Size: For Larege Dogs, Color: A Red, Size: For Larege Dogs
Great toy, heavy, good size, and durable for active chewers. My two dogs like to play tug a war a lot and this toy has held up fantastically! Afterwards they tend to chew toys to pieces but this one has stood the test of time so far. Great buy!
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Reviewed in the United States on May 14, 2026
S
Verified Purchase
SHUGRLTARA
Port Orchard, US
★★★★★ 4
Very tough & VERY heavy
Color: A Red, Size: For Larege Dogs
My dog is a half black lab/half pittie & a very aggressive chewer she goes through toy bones very quickly, to the point they're so slivered up within 2 days, sometimes slightly longer, I have to get rid of them, they never last long, except for whole antlers and some split antlers. Although she rarely touches the whole antlers . Anyways, this is a great tough toy for strong large breeds. It squeaks in the center if you squeeze hard enough but the toy/bone was a bit too heavy for my girl. I got her the next smaller one and she still plays with the big heavier one more than the one that fits her perfectly so go figure. It's a great product and definitely recommend for heavy chewers. I only took a star off because of the weight I was extremely surprised by how heavy it was, but that's not necessarily a bad thing depending on the dog, this size is just not for my dog. I think I've had it about a month and she does play with it, not as much as her other toys, but it's still in amazing shape even with her heavy chewing.
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Reviewed in the United States on April 21, 2024
A
Verified Purchase
Alannah Henry
Lowell, US
★★★★★ 5
Squeaky Toy for Aggressive Chewers
Color: A Violet, Size: For Larege Dogs
Size is great for our big dog, he loves to chew & squeak it. engagement, our dog loves playing fetch, hours of enjoyment.
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Reviewed in the United States on May 21, 2026
E
Verified Purchase
Elizabeth
Alexandria, US
★★★★★ 3
Maybe not for super aggressive chewers
Color: A Violet, Size: For Larege Dogs, Color: A Violet, Size: For Larege Dogs
I had high hopes for this toy, I was so excited about it. I have aggressive chewers, two Malinois/Dutch Shepard mixes and a Pit. The Shepard's chewed pieces off of it in literally 5 minutes. At the ends there is a small lip and that's where they started chewing pieces off. I had ordered a second one before I gave them this one (early Christmas present) because when I received it I noticed it felt very sturdy and durable. They do like it so since they already chewed this one I'm going to see if I can shave that lip off and maybe they won't get anymore pieces off. That'll decide if I return the one that arrives tomorrow. Maybe not good for super aggressive chewers. It's very thick and seems well made. Might just be better for other dogs.
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Reviewed in the United States on December 21, 2025

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