Anti Cardiolipin IgM ELISA Assay Kit

$340.00

The Eagle Biosciences Anti Cardiolipin IgM ELISA Assay Kit is an indirect solid phase immunoassay kit for the quantitative measurement of IgM class auto-antibodies directed against Cardiolipina-β2-glycoprotein complex in human serum or plasma. The Anti Cardiolipin IgM ELISA Assay Kit is intended for research use only and not intended for diagnostic procedures.

SKU: DCM112 Categories: , ,

Anti Cardiolipin IgM ELISA Assay Kit

For Research Use Only

Size: 1×96 wells
Sensitivity: 0.12 AU/mL
Dynamic Range: 5 – 80 AU/ml
Incubation Time: 2 hours
Sample Type: Serum, Plasma
Sample Size: 10 µl

Controls Included

Product Developed and Manufactured in Italy by Diametra

Additional Information

Assay Background

The first study on the anti-phospholipid antibodies began in 1906 when Wasserman introduced a serological test for syphilis. In 1942 it was discovered that the active component is a phospholipid called Cardiolipina. In the 1950’s it was observed that a large number of people appeared to be positive for syphilis tests but did not show any evidence of disease. Initially, the phenomenon was classified as a series of false positive syphilis tests, before a more accurate analysis revealed, for this group of patients, a high prevalence of autoimmune disorders including SLE and Sjögrens syndrome.
The term lupus anticoagulant (LA), used for the first time in 1972, derives from experimental observations in which an increased risk of thrombosis was observed, paradoxically, with the presence of some anticoagulants factors; the term LA is not totally correct, in fact the disease is present more frequently in patients without lupus and it is associated with thrombosis rather than to abnormal bleeding.

Some years later the role of a cofactor was investigated, the β2-glycoprotein I (apolipoprotein H) also called β2GPI, and its interactions with anionic phospholipids in human serum / plasma. This cofactor is a β-globulin with a molecular weight of 50 kDa that has a concentration of 200 μg / mL in plasma. The β2GPI is involved in the regulation of blood coagulation, inhibiting the intrinsic way. β2GPI in vivo is associated with negatively charged substances such as anionic phospholipids, heparin and lipoproteins. The region that binds phospholipids is in its fifth domain.

The acronym “aPL” (anti-phospholipid antibodies) indicates improperly antibodies directed against negatively charged phospholipids like Cardiolipina (CL), Phosphatidyl serine (PS) Phosphatidyl inositol (PI) and phosphatidic acid (PA); more correctly the term anti-phospholipid antibodies indicates those antibodies directed against the complex between β2GPI and anionic phospholipids that can bind to the fifth domain of β2GPI. Among these, the Cardiolipina is the most commonly used phospholipid as an antigen for determining the aPL by ELISA method. Diagnostic laboratories measure the antibodies directed against the complex between β2GPI and negatively charged phospholipids, as Phosphatidyl serine (PS) Phosphatidyl inositol (PI) and phosphatidic acid (PA). Some researchers suggest the use of PS instead of Cardiolipina in ELISA assays, for a more precise diagnosis. However, these antibodies against phospholipids are less commonly used, even if their use may increase the clinical sensitivity of patient samples with suspected

Assay Principle

Anti Cardiolipin IgM ELISA Assay Kit is based on the binding of antibodies directed against the antigenic complex between Cardiolipina and β2-Glycoprotein on human serum or plasma; this complex is coated on the microplate. In the first step, the antibodies present in calibrators, controls or prediluted samples bind to this antigen. After 60 minutes of incubation, the microplate is washed with a wash buffer to remove the non-reactive serum or plasma components. Then an anti-human IgM horseradish peroxidase conjugated solution recognizes the IgM class antibodies bound to the immobilized antigens. After 60 minutes of incubation, any excess enzyme conjugate, which is not specifically bound, is washed away with the wash buffer. Finally, a chromogenic substrate solution containing TMB is dispensed into the wells. After a 15 minute incubation, the color development is stopped by adding the stop solution. The solution turns yellow at this point. The level of color is directly proportional to the concentration of IgM antibodies present in the original sample. The concentration of IgM antibodies in the sample is calculated through a calibration curve.

Manual

Product Manual