MASP-2 is a MBL associated serine protease. Mannan-binding lectin (MBL) and ficolins, in complex with MASPs, are capable of activating the complement system, thus mediating the destruction of infectious agents. After oligomerisation of MBL (or L-Ficolin) MASP-2 will be bound and activated. Activated MASP-2 cleaves C4 and C2 and is crucial for the activation of downstream complement components. MASP-2 deficiency will result in an increased susceptibility for infections (e.g. leukemia patients in chemotherapy are at high risk of serious infections). Even more so then MBL deficiency, because MASP-2 is also required for complement activation by H- and L-ficolin. The ELISA detects MASP-2 in serum and plasma. The MASP-2 levels in plasma from healthy individuals range from 170 to 1196 ng/ml.
The human MASP-2 ELISA kit is to be used for the in vitro quantitative determination of human MASP-2 in serum, plasma and cell culture supernatant samples.
The human MASP-2 ELISA is a ready-to-use solid-phase enzyme-linked immunosorbent assay based on the sandwich principle which a working time of 3½ hours.
The efficient format of 2 plates with twelve disposable 8-well strips allow free choice of batch size for the assay.
Samples and standards are captured by a solid bound specific antibody.
Biotinylated tracer antibody will bind to captured human MASP-2.
Streptavidin-peroxidase conjugate will bind to the biotinylated tracer antibody.
Streptavidin-peroxidase conjugate will react with the substrate, tetramethylbenzidine (TMB).
The enzyme reaction is stopped by the addition of oxalic acid.
The absorbance at 450 nm is measured with a spectrophotometer. A standard curve is obtained by plotting the absorbance (linear) versus the corresponding concentrations of the human MASP-2 standards (log).
The human MASP-2 concentration of samples, which are run concurrently with the standards, can be determined from the standard curve.
The linearity of the assay was determined by serially diluting a sample containing 328 ng/ml human MASP-2. The diluted samples were measured in the assay. The line obtained a slope of 1.01 and a correlation coefficient of 0.999.