Human t-Plasminogen Activator/tPA ELISA Kit - Quantikine
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Summary
Precision
Recovery
Linearity
Data Examples
Product Datasheets
Preparation and Storage
Related Research Areas
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.
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.
⚠ 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 50 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.
50 µL Assay Diluent
50 µL Standard, Control, or Sample
200 µL Conjugate
200 µL Substrate Solution
50 µL Stop Solution
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.
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Citations: Showing 1 - 7
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Elevated Plasma Levels of Mature Brain-Derived Neurotrophic Factor in Major Depressive Disorder Patients with Higher Suicidal Ideation
Authors: Li, H;Zhao, M;Jiang, C;Zhao, H;Wu, C;Li, Y;Zhang, S;Xu, P;Mou, T;Xu, Y;Huang, M;
Brain sciences
Species: Human
Sample Types: Plasma
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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
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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
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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
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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
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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
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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
