Anti Cardiolipin Screen ELISA Assay Kit

$340.00

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

SKU: DCM144 Categories: , ,

Anti Cardiolipin Screen ELISA Assay Kit

For Research Use Only

Size: 1×96 wells
Sensitivity: 0.08 AU/mL
Dynamic Range: 5 – 80 AU/ml
Incubation Time: 2.5 hour
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 Cardiolipin. 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 Cardiolipin (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 Cardiolipin 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 Cardiolipin 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


Eagle Biosciences Anti Cardiolipin Screen ELISA Assay Kit allows to determine the unknown concentration of auto-antibodies directed against the Cardiolipin-β2-glycoprotein complex through two different calibration curve (one specific for IgG test, one specific for IgM test), two different conjugates linked to horseradish peroxidase (one specific for IgG test, the other specific for IgM test) and one microplate only. The principle of the method and the procedure are the same in both the tests. Use reagents for IgG or reagents for IgM depending on the isotype which is under investigation.

Anti Cardiolipin Screen ELISA Assay Kit is based on the binding of antibodies directed against the antigenic complex between Cardiolipin and β2-Glycoprotein on human serum or plasma; this complex is coated on the microplate. Through the first incubation of 60 minutes, the IgG and IgM class antibodies directed against these complexes and present in calibrators, controls, and prediluted samples bind to the antigenic complexes coated on the microplate. At the end of incubation, the microplate is washed with a Wash Solution to remove the non-reactive serum or plasma components.

Then an incubation with the conjugate is performed: in this step, the anti human IgG antibodies (Conjugate IgG, reactive 3) or anti human IgM (Conjugate IgM, reactive 6) recognize the IgG or IgM class antibodies (respectively) bound to the immobilized antigenic complexes. After a 60 minutes incubation, the excess of unbound conjugate is washed away with the Wash Solution. A chromogenic substrate solution (TMB Substrate) containing tetramethylbenzidine is then dispensed into the wells. After a 15 minutes incubation, the color development is stopped by adding the Stop Solution. The solution turns into yellow and the intensity of color is directly proportional to the concentration of IgG or IgM antibodies present in the sample. The concentration of IgG or IgM class antibodies directed against the Cardiolipin-β2-glycoprotein complex in the sample is derived from a calibration curve, specific for IgG or IgM.

Manual

Product Manual