Overview

  • Product name

    Mouse uPA ELISA Kit
    See all uPA kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    serum 8 2.8%
    Inter-assay
    Sample n Mean SD CV%
    serum 3 3.5%
  • Sample type

    Urine, Serum, Cell culture media, Hep Plasma, EDTA Plasma, Cit plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    31.7 pg/ml
  • Range

    62.5 pg/ml - 4000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Urine 112 101% - 124%
    Serum 109 103% - 118%
    Cell culture media 95 91% - 99%
    Hep Plasma 110 105% - 114%
    EDTA Plasma 113 108% - 120%
    Cit plasma 106 104% - 106%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse
    Does not react with: Cow
  • Product overview

    uPA in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of uPA protein in mouse serum, plasma, urine, and cell culture media.


    The 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. TMB Development Solution is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.


    Urokinase-type plasminogen activator (uPA) is a serine protease encoded by the PLAU gene and known to specifically cleaves the zymogen plasminogen to form the active enzyme plasmin. Activation of plasmin triggers a proteolytic cascade that participates in thrombolysis or extracellular matrix degradation depending on the physiological environment. This cascade has been involved in vascular diseases and cancer progression, thus making uPA an attractive drug target. This kit was raised against the alpha chain of uPA and calibrated to the full length protein.


     

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Mouse uPA Capture Antibody 1 x 600µl
    10X Mouse uPA Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent 4BR 1 x 6ml
    Mouse uPA Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • Relevance

    Urokinase type plasminogen activator (uPA) is secreted from cells as a precursor which is activated to the two chain form consisting of an A and B chain. The active high MW form is further processed by removal of an amino terminal fragment to an active low MW form (35 kDa). uPA is a serine protease that activates plasminogen toplasmin. High levels of uPA and plasminogen activator inhibitor type 1 (PAI 1) in breast cancer tissue extracts have been associated with rapid disease progression. The malignant phenotype of prostatic tumor cells correlates with the expression of both uPA and its cell membrane receptor (uPAR).
  • Alternative names

    • ATF
    • BDPLT5
    • EC 3.4.21.73
    • PLAU
    • QPD
    • u PA
    • U plasminogen activator
    • u-PA
    • UPA
    • UPAM
    • Urinary plasminogen activator
    • URK
    • Urokinase plasminogen activator
    • Urokinase type plasminogen activator
    see all
  • Database links

Associated products

Applications

Our Abpromise guarantee covers the use of ab245727 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • The uPA standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
  • The concentrations of uPA were measured in duplicates, interpolated from the uPA standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 50%, plasma (citrate) 50%, plasma (EDTA) 25%, and plasma (heparin) 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean uPA concentration was determined to be 7166 pg/mL in serum, 6222 pg/mL in plasma (citrate) and 7060 pg/mL in plasma (EDTA), and 4907 pg/mL in plasma (heparin).
  • The concentrations of uPA were measured in duplicates, interpolated from the uPA standard curves and corrected for sample dilution. Undiluted samples are as follows: urine 1.5%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean uPA concentration was determined to be 272.6 ng/mL in urine.
  • The concentrations of uPA were measured in duplicates, interpolated from the uPA standard curves and corrected for sample dilution. Undiluted samples are as follows: cell culture media 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).

Protocols

References

ab245727 has not yet been referenced specifically in any publications.

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