ALPHA 2 MACROGLOBULIN ELISA KIT
INTENDED USE
The Alpha 2 Macroglobulin ELISA Kit Assay is intended for the quantitative determination of α 2-Macroglobulin in urine, serum and plasma. For in vitro diagnostic use only.
SUMMARY AND EXPLANATION OF THE TEST
Alpha 2-Macroglobulin is one of the biggest plasma proteins with a molecular weight of 650-900kDa, dependent on the degree of glycosylation. It consists of 4 subunits. Alpha 2-Macroglobulin functions as protease inhibitor of all known classes of endopeptidase (serine-, cysteine-, aspartatand metalloproteases).
The measurement of urinary proteins in the urine allows the diagnosis of proteinuria. In case of a microalbuminuria (selective proteinuria) increased levels ( > 150 mg protein/24h) of albumin and transferrin (marker proteins of the tubulus) could be measured. In case of severe diseases of the kidney with pathological changes of the basal membran in the glomerulus (Chronic Glomerulonephritis, Tubulopathy, Nephrotic Syndrom) the protein level will rise up to > 3 g/24h and also the proteintyp will change to high molecular proteins like β-Lipoproteins and α 2-Macroglobulin (non-selective proteinuria).
Additionally previous investigations have shown that the determination of the two acute-phase proteins in the urins, C-reactive protein (CRP) and α 2-Macroglobulin, allows a noninvasive diagnosis of acute renal transplant dysfunction and rejection.
PRINCIPLE OF THE TEST
Alpha 2 Macroglobulin ELISA Kit
The Alpha 2 Macroglobulin ELISA Kit Assay is intended for the quantitative determination of α 2-Macroglobulin in urine, serum and plasma. In a first incubation step, the α 2-Macroglobulin in the samples is bound to polyclonal rabbit antibodies (in excess), which are immobilized to the microtiter wells. To remove all unbound substances, a washing step is carried out. In a second incubation step, a Peroxidase-labeled anti α2-Macroglobulin (POD-Antibody) antibody is added. After another washing step, to remove all unbound substances, the solid phase is incubated with the substrate, tetramethylbenzidine (TMB). An acidic stop solution is then added to stop the reaction. The color converts from blue to yellow. The intensity of the yellow color is directly proportional to the concentration of α 2-Macroglobulin in the sample. A dose response curve of the absorbance unit (optical density, OD) vs. concentration is generated, using the results obtained from the calibrators. α 2-Macroglobulin, present in the patient samples, is determined directly from this curve.
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