Human u-Plasminogen Activator/Urokinase Quantikine ELISA
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Summary
Precision
Recovery
Linearity
Data Examples
Product Datasheets
Preparation and Storage
Background
Related Research Areas
Human u-Plasminogen Activator/Urokinase Quantikine ELISA Summary
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Cell Lysates (50 uL), Serum (17 uL), EDTA Plasma (17 uL), Heparin Plasma (17 uL), Urine (10 uL)
Sensitivity
4.17 pg/mL
Assay Range
31.3 - 2,000 pg/mL (Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma, Urine)
Specificity
Natural and recombinant human uPA. This assay also recognizes uPA complexed with PAI-1 or uPAR.
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.
Product Summary
The Quantikine Human u-Plasminogen Activator/Urokinase Immunoassay is a 4.5 hour solid phase ELISA designed to measure uPA in cell culture supernates, cell lysates, serum, plasma, and urine. It contains NS0-expressed recombinant human uPA and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate recombinant human uPA. Results obtained using natural human uPA showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural human uPA.
Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision
Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma, Urine
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 230 | 629 | 1219 | 214 | 614 | 1188 |
| Standard Deviation | 4.8 | 8.9 | 29.3 | 15.1 | 39.8 | 82 |
| CV% | 2.1 | 1.4 | 2.4 | 7.1 | 6.5 | 6.9 |
Recovery
The recovery of uPA spiked to three different levels throughout the range of the assay was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=8) | 102 | 93-115 |
| Cell Lysates (n=4) | 97 | 81-116 |
| EDTA Plasma (n=4) | 87 | 83-94 |
| Heparin Plasma (n=4) | 89 | 81-110 |
| Serum (n=4) | 87 | 81-93 |
| Urine (n=4) | 98 | 81-119 |
Linearity
To assess the linearity of the assay, samples containing high concentrations of uPA were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay. Samples were diluted prior to assay.
Scientific Data
Product Datasheets
Preparation and Storage
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: u-Plasminogen Activator (uPA)/Urokinase
u-Plasminogen Activator (uPA) is a serine protease that converts plasminogen to plasmin, with roles in a variety of normal and pathological processes that include cell migration and tissue destruction. uPA is a potent marker of invasion and metastasis in a variety of human cancers including breast, stomach, colon, bladder, ovarian, brain, and endometrium.
Long Name:
Urokinase-type Plasminogen Activator
Entrez Gene IDs:
5328 (Human); 18792 (Mouse); 102135886 (Cynomolgus Monkey)
Alternate Names:
ATF; EC 3.4.21; EC 3.4.21.73; plasminogen activator, urokinase; PLAU; uPA; u-PA; uPlasminogen Activator; u-Plasminogen Activator; urinary; Urokinase; urokinase-type plasminogen activator
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.
Assay Procedure
Refer to the product Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
- Prepare all reagents, standard dilutions, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
- Add 100 µL of Assay Diluent to each well.
- Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
- Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
- Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
- Aspirate and wash 4 times.
- Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
- Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
100 µL Assay Diluent
50 µL Standard, Control, or Sample
200 µL Conjugate
200 µL Substrate Solution
50 µL Stop Solution
Citations for Human u-Plasminogen Activator/Urokinase Quantikine ELISA
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Citations: Showing 1 - 5
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Targeting uPA-uPAR interaction to improve intestinal epithelial barrier integrity in inflammatory bowel disease
Authors: Y Cheng, TR Hall, X Xu, I Yung, D Souza, J Zheng, F Schiele, M Hoffmann, ML Mbow, JP Garnett, J Li
EBioMedicine, 2021-12-18;75(0):103758.
Species: Human
Sample Types: Cell Culture Supernates
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CD276 is an important player in macrophage recruitment into the tumor and an upstream regulator for PAI-1
Authors: S Durlanik, K Fundel-Cle, C Viollet, HJ Huber, M Lenter, K Kitt, S Pflanz
Scientific Reports, 2021-07-21;11(1):14849.
Species: Human
Sample Types: Cell Culture Supernates
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p-Cresol affects reactive oxygen species generation, cell cycle arrest, cytotoxicity and inflammation/atherosclerosis-related modulators production in endothelial cells and mononuclear cells.
Authors: Chang M, Chang H, Chan C, Yeung S, Hsien H, Lin B, Yeh C, Tseng W, Tseng S, Jeng J
PLoS ONE, 2014-12-17;9(12):e114446.
Species: Human
Sample Types: Whole Cells
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Collective migration of cancer-associated fibroblasts is enhanced by overexpression of tight junction-associated proteins claudin-11 and occludin.
Authors: Karagiannis G, Schaeffer D, Cho C, Musrap N, Saraon P, Batruch I, Grin A, Mitrovic B, Kirsch R, Riddell R, Diamandis E
Mol Oncol, 2013-11-08;8(2):178-95.
Species: Human
Sample Types: Cell Culture Supernates
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Paracrine and autocrine signals induce and maintain mesenchymal and stem cell states in the breast.
Authors: Scheel C, Eaton E, Li S, Chaffer C, Reinhardt F, Kah K, Bell G, Guo W, Rubin J, Richardson A, Weinberg R
Cell, 2011-06-10;145(6):926-40.
Species: Human
Sample Types: Cell Culture Supernates
FAQs
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Human u-Plasminogen Activator/Urokinase Quantikine ELISA
By Jenny Roska
on 06/18/2022
Sample Tested: HEK293 human embryonic kidney cell line
