Cat. No.
MAK-0186
Product Name
Rat α-2-Macroglobulin
Storage
4°C
Species
Rat
Introduction
Alpha-2-macroglobulin (A2M) is an acute phase protein that is elevated in serum or plasma because of injury, infection or disease.It functions as an inhibitor of proteases such as trypsin and is believed to provide a protective role during the acute phase response.
Principle Of The Assay
The assay uses affinity purified rat A2M antibodies for solid phase (microtiter wells) immobilization and horseradish peroxidase (HRP) conjugated rat A2M antibodies for detection. Samples are first mixed with trypsin and incubated for one hour at room temperature. The trypsin reaction is then stopped by addition of trypsin inhibitor. Trypsin treatment normalizes reactivity of the A2M isoforms in nonacute phase and acute phase serum. Standards and diluted samples are then incubated in the microtiter wells for 45 minutes. The wells are subsequently washed. HRP conjugate is added and incubated for 45 minutes. This results in A2M molecules being sandwiched between the immobilization and detection antibodies. The wells are then washed to remove unbound HRP-conjugate. TMB is added and incubated for 20 minutes. If A2M is present a blue color develops. Color development is stopped by the addition of Stop solution, changing the color to yellow, and absorbance is measured at 450 nm. The concentration of A2M is proportional to absorbance and is derived from a standard curve.
