Mouse Alpha 2-Antiplasmin (Alpha2-AP) Active ELISA Assay

$870.00

The Eagle Biosciences Mouse Alpha 2-Antiplasmin (Alpha2-AP) Active ELISA Assay kit is for the quantitative determination of Active Alpha 2-Antiplasmin (Alpha2-AP) in mouse plasma by a microplate enzyme immunoassay (ELISA).

Mouse Alpha 2-Antiplasmin (Alpha2-AP) Active ELISA Assay

For Research Use Only

Size: 1×96 wells
Sensitivity: 0.025 ng/ml
Dynamic Range: 0.025 – 10 ng/ml
Incubation Time: 1.5 hours
Sample Type: Plasma
Sample Size: 100 µl

Product manufactured in the USA

Additional Information

Reference Values (Normal)

It has been determined in laboratory testing that mouse plasma contains approximately 300 µg/ml Alpha-2-antiplasmin.
It is suggested that each laboratory establish its own normal ranges.

Assay Background

This Mouse Alpha-2-antiplasmin (Alpha2-AP) Active ELISA Assay kit is for the quantitative determination of active a-2-antiplasmin in mouse plasma.  Alpha 2 antiplasmin (Alpha2-AP) is the major circulating inhibitor of plasmin, and it has a role in the regulation of intravascular fibrinolysis. Decreased levels of Alpha2 antiplasmin may play an important role in the increased capacity of the fibrinolytic function and may be beneficial in the treatment of thrombotic diseases, acute pulmonary embolism, and hepatic repair.

Assay Principle

Functionally active Alpha-2-antiplasmin present in plasma reacts with plasmin that has been coated and dried on a microtiter plate. Latent or complexed a-2-antiplasmin will not bind to the plate or be detected. Unbound a-2-antiplasmin is removed by washing and an anti-a-2-antiplasmin primary antibody is added. Excess primary antibody is removed by washing. The bound antibody, which is proportional to the original active a-2-antiplasmin present in the samples, is then reacted with the horseradish peroxidase-conjugated secondary antibody. Following an additional washing step, TMB substrate solution is then used for color development at 450 nm. The amount of color development is directly proportional to the concentration of active Alpha 2-antiplasmin in the sample.

  1. Add 100 µl of the Standards and unknowns to the wells in duplicate. Shake the plate at 300 rpm for 30 minutes at room temperature (RT). For a suggested plate layout, see Scheme I below.
  2. Wash the plate 3 times according to the following wash procedure:
    1. Remove the contents of each well by inversion of the plate.
    2. Tap out the remaining contents of the plate onto a lint free paper towel.
    3. Add 300 µL of 1x Wash Buffer.
    4. Let stand for 2 minutes.
    5. Repeat procedure two more times, then proceed to step “f”.
    6. Remove the contents of each well by inversion of the plate into an appropriate disposal device.
    7. Tap out the remaining contents of the plate onto a lint-free paper towel, then proceed to step 3.
  3. Add 100 µl 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 µl of the Secondary Antibody to each well. Shake the plate at 300rpm 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 10-20 minutes at RT.
  8. Stop the reaction by adding 50 µl of 1N H2SO4 to each well and read the plate at 450 nm.

Manual

Product Manual