Anti Beta 2 Glycoprotein 1 Screen ELISA Assay Kit

$300.00

The Eagle Biosciences Glycoprotein 1 Screen ELISA Assay Kit is an indirect enzyme-linked immunosorbent assay (ELISA) kit designed for the quantitative measurement of IgG or IgM class antibodies directed against the β2-Glycoprotein 1 in human serum or plasma. The Anti beta 2 Glycoprotein 1 Screen ELISA Assay Kit is intended for research use only and is not intended for diagnostic procedures.

SKU: DCM145 Categories: , ,

Anti Beta 2 Glycoprotein 1 Screen ELISA Assay Kit

For Research Use Only

Size: 1×96 wells
Sensitivity: 0.47 AU/mL
Dynamic Range: 10 – 160 AU/ml
Incubation Time: 2 hours
Sample Type: Serum, Plasma
Sample Size: 100 µL

Controls Included

Product Developed and Manufactured in Italy by Diametra

Additional Information

Assay Background


The antiphospholipid syndrome (APS) is a disorder that presents peculiar symptoms: arterial and venous thrombosis, thrombocytopenia, ulcers of the lower limbs, hemolytic anemia, loss of the fetus during pregnancy and is associated with the presence of antiphospholipid antibodies. Antiphospholipid antibodies represent a large and heterogeneous immunoglobulins group, including anticardiolipin antibodies and lupus anticoagulant. The former are diagnosed by on their reactivity with cardiolipin or other anionic phospholipids in ELISA test, while the latter is detected in phospholipid-dependent coagulation tests (aPTT, KCT, dRVVT).

In the early ’90s it was observed that antiphospholipid antibodies are not directed against anionic phospholipids, as long it was considered, but they react with plasma proteins bound to anionic (phospholipidic) surfaces. In fact about cardiolipin, it was observed the need for a cofactor for antibodies binding, this cofactor was identified in β2 glycoprotein 1 (β2-GP1). The β2 glycoprotein 1 is a plasma glycoprotein of molecular weight of 50 kD, which is complexed to lipoproteins for 40%. A patient with clinical suspicion of APS, who has a high titre of anti-beta2GP1 antibodies, even if  LAC or anti-cardiolipin antibodies are negative,  has a strong chance of having the antiphospholipid syndrome, because these antibodies recognize β2-GP1 bound to the surface of phospholipid (cardiolipin). These proteins seem to express their antigenicity only after contact with specific areas such as the anionic phospholipid surface or very hydrophilic plastic surfaces. The importance of ELISA anti-phospholipids test, including the anti-β2-GP1 test, lies in the fact that a positive test if associated with one or more of the symptoms can confirm a diagnosis of APS.

Assay Principle


Eagle Biosciences Anti beta 2 Glycoprotein 1 Screen test allows to determine the unknown concentration of auto-antibodies directed against the β2-Glycoprotein 1  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.

Glycoprotein 1 Screen ELISA Assay Kit is based on the initial binding of antibodies present in calibrators, controls or pre-diluted samples to the beta 2 Glycoprotein 1 coated on the inner surface of the microplate wells. After 60 minutes of incubation, the microplate is washed with a wash buffer to remove the non-reactive serum components. Then an anti-human IgG (Conjugate IgG, reactive 3) or IgM (Conjugate IgM, reactive 6) horseradish peroxidase conjugated solution recognizes the IgG or IgM (respectively) class antibodies bound to the immobilized antigens.

After 30 minutes incubation the excess of enzyme conjugate, which is not specifically bound, is washed away with a wash buffer. Finally, a chromogenic substrate solution containing TMB is dispensed into the wells. After 15 minutes of incubation, 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 IgG or IgM antibodies present in the original sample. The concentration of IgG or IgM antibodies present in the sample is calculated through a calibration curve.

Manual

Product Manual