Cardiovascular and Oxidative Stress Assay Kit

Rat tPA Total ELISA Assay

$1,130.00

The Eagle Biosciences Rat Tissue-type Plasminogen Activator (tPA) Total Antigen ELISA Assay kit is for the quantitative determination of total tPA in rat biological fluids by a ELISA assay (Enzyme-Linked Immunosorbent Assay).

SKU: TPN29-K01 Categories: , , ,

Rat tPA Total ELISA Assay

The Rat tPA Total ELISA Assay is For Research Use Only

Size: 1 x 96 wells
Sensitivity: 0.1 ng/ml
Standard Range: 0.1 – 50 ng/ml
Incubation Time: 2 hours
Sample Type: Biological Fluids
Sample Size: 100 µl
Alternative Name: Tissue-type Plasminogen Activator

Assay Principle

This Rat Tissue-type Plasminogen Activator (tPA) Total Antigen ELISA Assay Kit (Enzyme-Linked Immunosorbent Assay) is for the quantitative analysis of tPa levels in biological fluid.  This test kit operates on the basis of sandwich ELISA where free, latent and complexed tPa is quantified with the use of an HRP labeled secondary antibody.  The various forms of tPA present in the standard or unknown is captured by the tPA capture antibody coated on the well.  A primary antibody specific for tPA is then added to each well followed by the HRP conjugated secondary antibody.  The bound conjugated secondary antibody is detected by the addition of substrate, which generates an optimal color after 10 minutes. Quantitative test results may be obtained by the measure and comparison of the sample and standard absorbance readings when read with a microplate reader at 450 nm.

Related Products

Rat Tissue-type Plasminogen Activator (tPA) Active ELISA Assay Kit
Human tPA Total ELISA Assay
Mouse Tissue type Plasminogen Activator (tPA) Total ELISA Assay Kit

Additional Information

Assay Background

Tissue-type Plasminogen Activator (tPA) is a member of the serine proteinase family.  tPA functions to lyse fibrin clots into soluble plasmin fragments.  tPA is active in two forms, single chain and two chain.  The two-chain tPA is created via interaction with the plasmin product cleaving the single chain.  This two-chain form is regarded as the more active form.  Both single chain and two-chain tPA are complexable with PAI-1.  PAI-1 acts as an inhibitor for tPA by binding to the tPA and thus stifling its ability to lyse fibrin.  tPA can serve as an indicator of both myocardial infarction for patients with impaired fibrinolytic systems as well as a marker for type-II diabetes.

Assay Procedure

  1. Add 100 µl of the Standards and unknowns to the wells in duplicate.  For a suggested plate layout, see Scheme I below.  Shake the plate at 300 rpm for 30 minutes at RT.
  2. Wash the plate 3 times according to the following wash procedure:
    • Remove the contents of each well by inversion of the plate.
    • Add 300 mL of 1x Wash Buffer to each well.
    • Let stand for 2-3 minutes.
    • Repeat procedure two more times, then proceed to next step.
    • Remove the contents of each well by inversion of plate into an appropriate disposal device.
    • Tap out the remaining contents of the plate onto a lint free paper towel, then proceed to step 3.
  3. Add 100 ml of the Primary Antibody to each well.  Shake the plate at 300 rpm for 30 minutes at RT.
  4. Wash the plate three times as in step 2.
  5. Add 100 ml of the Secondary Antibody to each well. Shake the plate at 300 rpm for 30 minutes at RT.
  6. Wash the plate three times as in step 2.
  7. Add 100 µl of TMB Substrate to each well. Shake the plate at 300 rpm for 10 minutes at RT.
  8. Stop the reaction by adding 50 µl of 1N H2SO4 to each well and read the plate at 450 nm.

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