Human t-Plasminogen Activator/tPA Quantikine ELISA Kit

(1 citations)   
  • 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.
Control Available
QC207, Quantikine Immunoassay Control Set 864 for Human tPA - Please Inquire
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
Sample123123
n202020202020
Mean4981475287849014602898
Standard Deviation3648.488.832.162.3115
CV%7.23.33.16.64.34

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 Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: t-Plasminogen Activator/tPA
    • Long Name
      Tissue Plasminogen Activator
    • Entrez Gene IDs
      5327 (Human); 18791 (Mouse); 25692 (Rat);
    • Alternate Names
      Alteplase; DKFZp686I03148; EC 3.4.21; EC 3.4.21.68; plasminogen activator, tissue type; plasminogen activator, tissue; PLAT; Reteplase; tissue plasminogen activator (t-PA); tissue-type plasminogen activator; TPA; T-PA; tPlasminogen Activator; t-Plasminogen Activator;
    Related Research Areas
    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:

    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.

    1 Citations: Showing 1 - 1

    1. 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;7(1):6871.
      Species: Human
      Sample Type: Serum
    ELISA Controls
    Description Application Cat# Citations Images  

    Quantikine Immunoassay Control Set 864 for Human tPA

    Ctrl QC207
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    Supplemental ELISA Products
    Description Application Cat# Citations Images  

    Quantikine ELISA Wash Buffer 1

    ELISA WA126 6
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