DiaPharma Group, Inc.

The PAP complex test can be used to detect baseline as well as elevated levels of this complex which may occur during thrombotic events, in cases of endogenous hyperfibrinolysis and during thrombolytic therapy.

Test principle

The TC PAP-test is a solid phase enzyme immunoassay.

Specificity

The TC PAP ELISA measures plasmin-alpha2-antiplasmin complexes only. It is not affected by free plasminogen, a-2-antiplasmin or other plasmin complexes. The assay range is 15-150ng/mL for low PAP-levels and 15-1500 ng/mL for high PAP-levels. Normal plasma values will range from 0-514 ng/mL PAP complexes. The inter- and intra-assay variations are less than 10% and 5%, respectively.

Test samples

Use "inhibitor cocktail" plasma samples (citrate plasma containing 2000 KlU/mL aprotinin and 20mM benzamidine). For this purpose collect the blood of patients to be tested in precoated plastic or siliconized tubes already containing 3.2% buffered citrate in 1:10 ratio to blood as anticoagulant as well as aprotinin and benzami-dine in a ratio yielding the final concentrations of 2000 KJU/mL and 20mM, respectively. After filling, samples should be gently mixed by inverting the tube 5 times, and, then are placed in a crushed ice-water mixture. Centrifuge the blood within 30 min. after the puncture at 2000 x g for 30 min. at 4°C (preferably in a cold centrifuge with swing out rotor). Immediately after centrifugation, plasma should be carefully pipetted off and collected in a small plastic tube. After carefully mixing of the pooled plasma, precooled and prelabelled plastic tubes for Storage are filled. The freezing operation is very important: snap-freezing, that means almost instantaneous freezing, is highly recommended. Storage temperature should be kept constantly below a temperature of-30°C, and preferably at -70°C. The total time between blood collection and plasma freezing should not exceed 90 min. Thawing and refreezing of plasma aliquotes is not recommended. Thawing for assay is achieved rapidly using a waterbath at 37°C. After thawing, plasma samples are placed in a crushed ice-water mixture until analysis. Haemolytic and lipaemic plasmas may be used if no other samples are available.

Kit components

1) Coating antibody: 4 x lyophilized monoclonal anti PAP coating antibody (TC-Code BZ).

2) Peroxidase conjugated antibody: 5 x peroxidase conjugated anti-Plasminogen antibody (concentrated) (TC-Code BK).

3) Standard: 5 x lyophilized purified PAP complexes (TC-Code BM)