Mouse Plasminogen Total Antigen ELISA Assay Kit

$700.00

The Eagle Biosciences Mouse Plasminogen Total Antigen ELISA Assay Kit is intended for the quantitative determination of total Plasminogen antigen in mouse plasma, serum, urine, cell culture supernatant and tissue extracts. The Plasminogen Total Antigen ELISA Assay Kit is for research use only and should not be used for diagnostic procedures.

Mouse Plasminogen Total Antigen ELISA Assay Kit

For Research Use Only

Size: 1×96 wells
Dynamic Range: 2.5-500 ng/ml
Incubation Time: 1.5 hours
Sample Type: Mouse Plasma, Serum, Urine, Cell Culture Media, Tissue Extracts
Sample Size: 100 µL

Additional Information

Assay Background

Plasminogen is a single chain glycoprotein zymogen and is the precursor of the fibrinolytic enzyme plasmin. Plasminogen deficiencies are classified as hypoplasminogenemia (Type I) or dysplasminogenemia (Type II) and are associated with decreased extracellular fibrin clearance leading to mucous membrane lesions and ligneous conjunctivitis.

Assay Principle

The Eagle Biosciences Mouse Plasminogen Total Antigen ELISA Assay Kit is an Enzyme Immunoassay for the quantitative determination of total Plasminogen antigen in mouse serum, plasma, urine, cell culture supernatant and tissue extracts. This assay employs the quantitative sandwich enzyme immunoassay technique. An antibody specific for Mouse Plasmin has been pre-coated onto a microtiter plate. Standards or samples are pipetted into the wells and any Plasminogen or plasmin antigen present is bound by the immobilized antibody. A polyclonal anti-mouse Plasminogen antibody is then added and bound to the captured proteins. After washing away any unbound substances, a secondary antibody conjugated to HRP is added and incubate. After washing steps are carried out, a substrate solution (TMB) is added to the wells and color develops in proportion to the amount of Plasminogen bound in the initial step. The color development is stopped by the addition of acid and the intensity of the color is measured at a wavelength of 450 nm ±2 nm. The concentration of Plasminogen in the sample is then determined by comparing the O.D of samples to the standard curve.

Assay Procedure

  1. Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal it.
  2. Add 100 µl standards and samples into each well. Incubate for 30 minutes at RT with shaking at 300 rpm.
  3. Aspirate each well and wash, repeating the process 4 times for a total 5 washes. Wash by filling each well with wash buffer (350 μl) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each is essential to good performance. After the last wash, remove any remaining distilled water by aspirating, decanting or blotting against clean paper towels.
  4. Add 100 µl anti-mouse Plasminogen antibody into each well. Incubate for 30 minutes at RT with shaking at 300 rpm.
  5. Wash as according to step 3.
  6. Add 100 µl Secondary antibody-conjugated HRP into each well. Incubate for 30 minutes at RT with shaking at 300 rpm.
  7. Wash as according to step 3.
  8. Add 100 μl of TMB Substrate to each well. Incubate for 2-7 minutes at room temperature in dark.
  9. Add 50 μl Stop solution into each well.
  10. Read the OD with a microplate reader at 450 nm immediately.

Typical Standard Curve

Manual

Product Manual