Human TPA ELISA Kit, Fluorescent is a single-wash 90-min Simplestep used to quantify Human TPA with a sensitivity of 7.27 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Fluorescent Sandwich ELISA - 530/570/590 nm readout : works on any standard plate reader
Images
Key facts
- Detection method
- Fluorescent
- Sample types
- Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma
- Assay type
- Sandwich (quantitative)
- Reactive species
- Human
- Range
- 9.77 - 20000 pg/mL
- Assay time
- 1h 30m
- Sensitivity
- = 7.27 pg/mL
Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application sELISA | Reactivity Reacts | Dilution info - | Notes - |
Target data
Function
Converts the abundant, but inactive, zymogen plasminogen to plasmin by hydrolyzing a single Arg-Val bond in plasminogen. By controlling plasmin-mediated proteolysis, it plays an important role in tissue remodeling and degradation, in cell migration and many other physiopathological events. During oocyte activation, plays a role in cortical granule reaction in the zona reaction, which contributes to the block to polyspermy (By similarity).
Alternative names
Tissue-type plasminogen activator, t-PA, t-plasminogen activator, tPA, PLAT
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Human TPA ELISA Kit, Fluorescent is a single-wash 90-min Simplestep used to quantify Human TPA with a sensitivity of 7.27 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Fluorescent Sandwich ELISA - 530/570/590 nm readout : works on any standard plate reader
Key facts
- Detection method
- Fluorescent
- Sample types
- Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma
- Assay type
- Sandwich (quantitative)
- Reactive species
- Human
- Range
- 9.77 - 20000 pg/mL
- Assay time
- 1h 30m
- Assay Platform
- Pre-coated microplate (12 x 8 well strips)
- Sensitivity
- = 7.27 pg/mL
Precision
Intra assay
| Sample | n | C.V. |
|---|---|---|
Sample Human Serum | n 5 | C.V. 3.7 |
Inter assay
| Sample | n | C.V. |
|---|---|---|
Sample Human Serum | n 3 | C.V. 10.5 |
Recovery
Sample specific recovery
| Sample type | Average % | Range |
|---|---|---|
Sample type Cell culture supernatant | Average % = 119 | Range 116 - 123 % |
Sample type Serum | Average % = 104 | Range 100 - 107 % |
Sample type EDTA Plasma | Average % = 84 | Range 82 - 86 % |
Sample type Heparin Plasma | Average % = 97 | Range 94 - 99 % |
Sample type Citrate plasma | Average % = 99 | Range 97 - 104 % |
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- +4°C
- Storage information
- +4°C
Notes
Tissue Plasminogen Activator (TPA) in vitro CatchPoint SimpleStep ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Tissue Plasminogen Activator (TPA) protein in human serum, plasma and cell culture supernatant.
This CatchPoint SimpleStep ELISA kit has been optimized for Molecular Devices Microplate Readers. Click here for a list of recommended Microplate Readers.
If using a Molecular Devices' plate reader supported by SoftMax® Pro software, a preconfigured protocol for these CatchPoint SimpleStep ELISA Kits is available with all the protocol and analysis settings at www.softmaxpro.org.
The CatchPoint SimpleStep ELISA employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. CatchPoint HRP Development Solution containing the Stoplight Red Substrate is added. During incubation, the substrate is catalyzed by HRP generating a fluorescent product. Signal is generated proportionally to the amount of bound analyte and the intensity is measured in a fluorescence plater reader at 530/570/590 nm Excitation/Cutoff/Emission.
Tissue Plasminogen Activator (Tissue-type plasminogen activator or TPA) is a circulating serine protease involved in the breakdown of clots. TPA converts inactive plasminogen to active plasmin; in turn plasmin degrades the fibrin matrix in clots. In addition, plasmin can cleave TPA at Arg-310 with results in a two chain disulphide linked TPA that has even greater proteolytic activity. TPA is synthesized in many tissues and is secreted into most extracellular body fluids. Recombinant TPA is used medically to resolve or prevent blood clots in ischemic stroke or myocardial infarction.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Tissue Plasminogen Activator (tPA) also known as a plasminogen activator acts mechanically to convert plasminogen to plasmin a serine protease. This reaction occurs at the surface of a fibrin clot leading to clot degradation a process known as fibrinolysis. tPA has a molecular mass of approximately 70 kDa. It is mainly expressed in vascular endothelial cells and is released into the bloodstream in response to stimuli such as circulatory stasis or endothelial damage.
- Biological function summary
TPA plays a critical role in thrombolysis by breaking down blood clots into their soluble components. It regulates plasminogen function by cleaving this zymogen to yield the active protease plasmin. This function makes tPA integral in maintaining hemostasis and it does not form a part of a larger protein complex. The activity and the regulation of tPA are important for preventing pathologic clotting which can lead to cardiovascular complications.
- Pathways
TPA is central to the fibrinolytic pathway. This pathway facilitates the conversion of plasminogen to plasmin enabling clot resolution. In addition tPA interacts with other proteins such as urokinase plasminogen activator (uPA) and inhibitors like plasminogen activator inhibitor-1 (PAI-1). The balance between tPA and its inhibitors is important for the regulation of fibrinolytic activity impacting hemostatic and thrombotic events.
- Associated diseases and disorders
TPA connects closely with conditions like stroke and myocardial infarction due to its thrombolytic properties. In ischemic stroke excessive or insufficient tPA activity can disrupt normal blood flow leading to tissue damage. Additionally in myocardial infarction tPA's role in breaking down clots proves important for restoring coronary blood flow. It is also linked with proteins like fibrinogen as they serve as substrates in the clot degradation process and with PAI-1 which modulates its activity and influences disease progression.
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5 product images
Sandwich ELISA - Human TPA ELISA Kit, Fluorescent (ab229408)
Example of human Tissue Plasminogen Activator (TPA) standard curve in Sample Diluent NS.
The Tissue Plasminogen Activator (TPA) standard curve was prepared as described in Section 10. Raw data generated on SpectraMax M4 Multi-Mode Microplate Reader is shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
Sandwich ELISA - Human TPA ELISA Kit, Fluorescent (ab229408)
Interpolated concentrations of TPA in human serum and plasma.
The concentrations of TPA were measured in duplicate and interpolated from the TPA standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean TPA concentration was determined to be 4,710 pg/mL in serum and 4,270 pg/mL in plasma (heparin).
Sandwich ELISA - Human TPA ELISA Kit, Fluorescent (ab229408)
Interpolated concentrations of TPA in human serum from 10 male donors.
Serum from 10 apparently healthy male donors was measured in duplicate. The mean TPA concentration was determined to be 2,741 pg/mL with a range of 1,836- 4,012 pg/mL in male donors.
Sandwich ELISA - Human TPA ELISA Kit, Fluorescent (ab229408)
Comparison of secreted TPA in unstimulated and PHA-stimulated human PBMC.
PBMC were grown in the absence or presence of phytohemagglutinin (PHA) for 2 days. TPA was measured in 4-fold diluted cell culture supernatants of unstimulated and PHA stimulated PBMC. Measured values were interpolated from the TPA Standard Curve diluted in Sample Diluent NS and DF corrected. Mean +/-SD, n=2, are graphed.
Sandwich ELISA - Human TPA ELISA Kit, Fluorescent (ab229408)
Titration of human serum and plasma (heparin) within the working range of the assay.
Background-subtracted data values (mean +/- SD, n =2) are graphed.
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