Human u-Plasminogen Activator/Urokinase Quantikine ELISA

R&D Systems | Catalog # DUPA00

R&D Systems
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Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Cell Lysates (50 µL), Serum (17 µL), EDTA Plasma (17 µL), Heparin Plasma (17 µL), Urine (10 µL)

Sensitivity

4.17 pg/mL

Assay Range

31.3-2000 pg/mL (Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma, Urine)
View all u-Plasminogen Activator (uPA)/Urokinase ELISA Kits »

Product Summary for Human u-Plasminogen Activator/Urokinase Quantikine ELISA

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.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Species

Human

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.

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, EDTA Plasma, Heparin Plasma, Serum, 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 82.0
CV% 2.1 1.4 2.4 7.1 6.5 6.9

Recovery for Human u-Plasminogen Activator/Urokinase Quantikine ELISA

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.

Human u-Plasminogen Activator (uPA)/Urokinase ELISA Linearity

Scientific Data Images for Human u-Plasminogen Activator/Urokinase Quantikine ELISA

Human u-Plasminogen Activator (uPA)/Urokinase ELISA Standard Curve

Human u-Plasminogen Activator (uPA)/Urokinase ELISA Standard Curve

Preparation and Storage

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Stability & 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

Alternate Names

PLAU, uPA, uPlasminogen Activator, Urokinase

Entrez Gene IDs

5328 (Human); 18792 (Mouse); 102135886 (Cynomolgus Monkey)

Gene Symbol

PLAU

Additional u-Plasminogen Activator (uPA)/Urokinase Products

Product Documents for Human u-Plasminogen Activator/Urokinase Quantikine ELISA

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human u-Plasminogen Activator/Urokinase Quantikine ELISA

For research use only

⚠ 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.

Citations for Human u-Plasminogen Activator/Urokinase Quantikine ELISA

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  • Human u-Plasminogen Activator/Urokinase Quantikine ELISA
    Name: Jenny Roska
    Sample Tested: HEK293 human embryonic kidney cell line
    Verified Customer | Posted 06/18/2022
    Human u-Plasminogen Activator/Urokinase Quantikine ELISA DUPA00

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Protocols

View specific protocols for Human u-Plasminogen Activator/Urokinase Quantikine ELISA (DUPA00):

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
    3. 100 µL Assay Diluent
  3.   Add 100 µL of Assay Diluent to each well.
    4. 50 µL Standard, Control, or Sample
  4.   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.
  5.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
    6. 200 µL Conjugate
  6.   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.
  7.   Aspirate and wash 4 times.
    8. 200 µL Substrate Solution
  8.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
    9. 50 µL Stop Solution
  9.   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.

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FAQs

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Associated Pathways

Blood Coagulation Signaling Pathways Blood Coagulation Signaling Pathway Thumbnail