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This AlphaLISA® immunoassay kit enables the isotyping of mouse IgM antibodies in different sample matrices, including cell culture media. The IgM subtype is the first antibody found during an immune response and is responsible for agglutination and cytolytic reactions.
There are five classes of mammalian immunoglobulins: IgA, IgD, IgE, IgM, and IgG. IgG is the most abundant immunoglobulin and is equally distributed in blood and tissue. In mice, the IgG class is further divided into four subclasses: IgG1, IgG2a/ IgG2c (strain specific), IgG2b, and IgG3. Serum IgM exists as a pentamer in mammals, predominates in primary immune responses to most antigens, is the most efficient complement fixing immunoglobulin and comprises approximately 10% of normal human serum Ig content. Each of the five monomers is composed of two light chains (either kappa or lambda) and two heavy chains. IgM can cause cell agglutination as a result of recognition of epitopes on invading microorganisms. This antibody-antigen immune complex is then destroyed by complement fixation or receptor mediated endocytosis by macrophages. Typically mouse serum and plasma samples contain about 0.5 to 6.5 mg/mL of IgM. The present kit permits detection of mouse IgM (i.e. analyte) in different sample matrices, including different cell culture media and monkey serum.
AlphaLISA technology allows the detection of molecules of interest in a no-wash, highly sensitive, quantitative assay. In an AlphaLISA assay, a biotinylated anti-analyte antibody binds to the Streptavidin-coated Donor beads while another anti-analyte antibody is conjugated to AlphaLISA Acceptor beads. In the presence of the analyte, the beads come into close proximity. The excitation of the Donor beads causes the release of singlet oxygen molecules that triggers a cascade of energy transfer in the Acceptor beads, resulting in a sharp peak of light emission at 615 nm.
Protein Analysis & Detection
|Assay Target Class