alpha 2-Macroglobulin (a2-M) ELISA Assay Kit


The alpha 2-Macroglobulin (a2-M) ELISA Assay Kit is intended for the quantitative determination of α2-macroglobulin in urine, serum and plasma. For research use only. Not for use in diagnostic procedures.

alpha 2-Macroglobulin a2-M ELISA Assay Kit

alpha 2-Macroglobulin a2-M ELISA Assay Kit manufactured in Germany by Immundiagnostik

Size: 1×96 wells
Standard Range: Plasma, serum: 1.3–3.0 g/l, Urine: < 0.18 mg/l; corresponds to 180 ng/ml
Incubation Time: 1h, 1h, 10-20min
Sample Type: Urine, Serum, Plasma
Sample Size: 100 µl (Urine); 50 µl (Serum, Plasma)
Alternative Names: Human alpha 2-Macroglobulin ELISA, Human a2-M ELISA, a2-M ELISA
For Research Use Only
Controls Included

Quality Control and Reference Range
It is recommended to use external controls for internal quality control, if possible. Control samples should be analysed with each run. Results, generated from the analysis of control samples, should be evaluated for acceptability using appropriate statistical methods. The results for the patient samples may not be valid if within the same assay one or more values of the quality control sample are outside the acceptable limits.

Scientific Background

Alpha-2-Macroglobulin (α2M) is one of the biggest plasma proteins, with a molecular weight of 650–900 kDa, depending on the degree of glycosylation. It consists of 4 identical subunits. α2M inhibits all known classes of endopeptidases by binding them and thereby blocking their active sites. The α2M-endopeptidase complex is then cleared rapidly from the circulation by the endocytotic proteinase clearance pathway. α2M also binds, transports and regulates many other molecules like defensins, myelin basic protein, and a host of other cytokines, growth factors, and hormones.
Measuring urinary proteins allows the diagnosis of proteinuria, which is defined as 150 mg protein/day. Proteinuria can be divided into prerenal, renal (glomerular or tubular), and postrenal proteinuria depending on the localisation of the kidney dam-age. Differential diagnosis can be achieved by measuring certain marker proteins of different molecular weights. Very large proteins, such as α2M, are completely restrict-ed from glomerular filtration in the kidneys. Thus, detecting α2M in urine is evidence of postrenal damage, when unfiltered serum proteins leak into the urine. Causes of postrenal damage are inflammation or hematuria as a consequence of renal stones or carcinomas.

Reference range
Plasma and serum: 1.3–3.0g/l
Urine: < 0.18mg/l; corresponds to 180ng/ml
We recommend each laboratory to establish its own reference range.

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Additional Information

Assay Principle

In a first incubation step, the α2-macroglobulin in the samples is bound to polyclonal rabbit antibodies (in excess), which are immobilised to the microtiter wells. To remo-ve all unbound substances, a washing step is carried out. In a second incubation step, a peroxidase-labelled anti α2-macroglobulin antibody (POD-antibody) is added. Af-ter another washing step to remove all unbound substances, the solid phase is incu-bated with the substrate, tetramethylbenzidine (TMB). An acidic stop solution is then added to stop the reaction. The colour converts from blue to yellow. The intensity of the yellow colour is directly proportional to the concentration of α2-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. α2-macroglobulin, present in the patient samples, is determined directly from this curve.

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