Human t-Plasminogen Activator/tPA Quantikine ELISA Kit

R&D Systems | Catalog # DTPA00

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), Serum (13 µL), EDTA Plasma (13 µL), Heparin Plasma (13 µL), Citrate Plasma (13 µL), Saliva (50 µL), Urine (25 µL), Human Milk (13 µL)

Sensitivity

16.1 pg/mL

Assay Range

78.1-5000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma, Saliva, Urine, Human Milk)
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Product Summary for Human t-Plasminogen Activator/tPA Quantikine ELISA Kit

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.

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 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.

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, Citrate Plasma, EDTA Plasma, Heparin Plasma, Human Milk, Saliva, 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) 498 1475 2878 490 1460 2898
Standard Deviation 36.0 48.4 88.8 32.1 62.3 115
CV% 7.2 3.3 3.1 6.6 4.3 4.0

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

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

Human t-Plasminogen Activator/tPA ELISA Standard Curve

Human t-Plasminogen Activator/tPA 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: t-Plasminogen Activator/tPA

tPA (tissue-type Plasminogen Activator) is a serine protease that is secreted by vascular endothelial cells, fibroblasts, neurons, microglia, and astrocytes. The partially active single chain can be further processed to full activity by Plasmin, Kallikrein 1, or Coagulation Factor Xa.  Active tPA converts Plasminogen to Plasmin, a fibrinolytic protease, by hydrolyzing an Arg-Val peptide bond in Plasminogen. Unusually high levels of tPA activity can result in excessive bleeding, and low levels of tPA activity can result in thrombosis or embolism. tPA-mediated breakdown of the extracellular is also involved in promoting synaptic plasticity, neurite outgrowth, and synapse remodeling. Human tPA contains an N-terminal fibronectin type-1 domain, an epidermal growth factor-like domain, two kringle domains, and a serine protease catalytic domain.

Long Name

Tissue Plasminogen Activator

Alternate Names

Alteplase, PLAT, Reteplase, T-PA, tPlasminogen Activator

Entrez Gene IDs

5327 (Human); 18791 (Mouse); 25692 (Rat)

Gene Symbol

PLAT

Additional t-Plasminogen Activator/tPA Products

Product Documents for Human t-Plasminogen Activator/tPA Quantikine ELISA Kit

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 t-Plasminogen Activator/tPA Quantikine ELISA Kit

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 t-Plasminogen Activator/tPA Quantikine ELISA Kit

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  • Human t-Plasminogen Activator/tPA Quantikine ELISA Kit
    Name: Leslie Priddy
    Sample Tested: Serum
    Verified Customer | Posted 04/01/2018

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Protocols

View specific protocols for Human t-Plasminogen Activator/tPA Quantikine ELISA Kit (DTPA00):

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
  3.   Add 50 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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