Human t-Plasminogen Activator/tPA ELISA Kit - Quantikine

Catalog # Availability Size / Price Qty
DTPA00
Control Products Available
Human t-Plasminogen Activator/tPA ELISA Standard Curve
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Product Details
Procedure
Citations (6)
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Human t-Plasminogen Activator/tPA Quantikine ELISA Kit 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), Serum (13 uL), EDTA Plasma (13 uL), Heparin Plasma (13 uL), Citrate Plasma (13 uL), Saliva (50 uL), Urine (25 uL), Human Milk (13 uL)
Sensitivity
16.1 pg/mL
Assay Range
78.1 - 5,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma, Saliva, Urine, Human Milk)
Specificity
Natural and recombinant human tPA. This assay also recognizes tPA complexed with PAI-1.
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 t-Plasminogen Activator/tPA Immunoassay is a 4.5 hour solid phase ELISA designed to measure tPA levels in cell culture supernates, serum, plasma, saliva, urine, and human milk. It contains CHO cell-expressed recombinant human tPA and antibodies raised against the recombinant protein. Results obtained for naturally occurring human tPA showed linear curves that were parallel to the standard curves obtained using the Quantikine Human tPA Immunoassay standards. These results indicate that this kit can be used to determine relative mass values for natural human tPA.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components

Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma, Saliva, Urine, Human Milk

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 498 1475 2878 490 1460 2898
Standard Deviation 36 48.4 88.8 32.1 62.3 115
CV% 7.2 3.3 3.1 6.6 4.3 4

Recovery

The recovery of human tPA spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 93 86-97
Citrate Plasma (n=4) 86 80-98
EDTA Plasma (n=4) 87 80-97
Heparin Plasma (n=4) 86 79-100
Serum (n=4) 95 87-109
Urine (n=4) 97 88-110

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human tPA were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human t-Plasminogen Activator/tPA ELISA Linearity

Scientific Data

Human t-Plasminogen Activator/tPA ELISA Standard Curve

Product Datasheets

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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.
⚠ 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 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. 50 µL Assay Diluent
  4.   Add 50 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   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.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 200 µL Conjugate
  9.   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.
  10.   Aspirate and wash 4 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  13. 50 µL Stop Solution
  14.   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.

Citations for Human t-Plasminogen Activator/tPA Quantikine ELISA Kit

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.

6 Citations: Showing 1 - 6
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  1. Oxidative stress-induced endothelial dysfunction and decreased vascular nitric oxide in COVID-19 patients
    Authors: V Montiel, I Lobysheva, L Gérard, M Vermeersch, D Perez-Morg, T Castelein, JB Mesland, P Hantson, C Collienne, D Gruson, MA van Dievoe, A Persu, C Beauloye, M Dechamps, L Belkhir, A Robert, M Derive, PF Laterre, AHJ Danser, X Wittebole, JL Balligand
    EBioMedicine, 2022-02-23;77(0):103893.
    Species: Human
    Sample Types: Serum
  2. The pleiotropic beneficial intervention of olive oil intake on the Alzheimer's disease onset via fibrinolytic system
    Authors: EE Tzekaki, M Tsolaki, &alphaA Pantazaki, G Geromichal, E Lazarou, M Kozori, Z Sinakos
    Experimental gerontology, 2021-04-06;150(0):111344.
    Species: Human
    Sample Types: Serum
  3. Effects of antithrombotic drugs on the prothrombotic state in patients with atrial fibrillation: The west Birmingham atrial fibrillation project
    Authors: C Voukalis, GYH Lip, E Shantsila
    Thrombosis Research, 2021-02-09;200(0):149-155.
    Species: Human
    Sample Types: Plasma
  4. Increased ratio of mature BDNF to precursor-BDNF in patients with major depressive disorder with severe anhedonia
    Authors: C Wu, J Lu, S Lu, M Huang, Y Xu
    J Psychiatr Res, 2020-05-12;126(0):92-97.
    Species: Human
    Sample Types: Plasma
  5. Combined serum levels of multiple proteins in tPA-BDNF pathway may aid the diagnosis of five mental disorders
    Authors: S Chen, H Jiang, Y Liu, Z Hou, Y Yue, Y Zhang, F Zhao, Z Xu, Y Li, X Mou, L Li, T Wang, J Zhao, C Han, Y Sui, M Wang, Z Yang, Y Lu, Y Zhu, J Li, X Shen, F Sun, Q Chen, H Chen, Y Yuan
    Sci Rep, 2017-07-31;7(1):6871.
    Species: Human
    Sample Types: Serum
  6. The serum protein levels of the tPA-BDNF pathway are implicated in depression and antidepressant treatment
    Authors: H Jiang, S Chen, C Li, N Lu, Y Yue, Y Yin, Y Zhang, X Zhi, D Zhang, Y Yuan
    Transl Psychiatry, 2017-04-04;7(4):e1079.
    Species: Human
    Sample Types: Serum

FAQs

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Human t-Plasminogen Activator/tPA Quantikine ELISA Kit
By Leslie Priddy on 04/01/2018
Sample Tested: Serum