ELISA, Western Blot. This IgY fraction antibody of anti-Human macrophage migration inhibitory factor (MIF) has been tested for use in ELISA and immunoblotting. Although not tested, this antibody may also be useful for neutralization assays, immunohistochemistry and flow cytometry. The antibody recognizes 12,300 MW mature human MIF. The MIF gene encodes a protein of 115 aa. The initiating methionine is cleaved leaving a mature protein of 114 aa. Reactivity in other immunoassays is unknown.
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Description
This product is an IgY fraction antibody purified from monospecific chicken egg yolks by a multi-step process which includes selective precipitation and salt fractionation followed by extensive dialysis against the buffer stated above. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Chicken Serum. The antibody is directed against the 12,300 MW human MIF protein and is useful in determining its presence in various assays. If neutralization experiments are performed for human MIF activity in bioassays, it is recommended to incubate the sample with a 1:500 dilution of the antibody for at least 4 hours before being tested. A control of similarly diluted normal chicken IgG is recommended. If FACS analysis experiments are performed for human MIF caution should be exhibited as the F( c) domain of the chicken IgG molecule may interact with cells non-specifically.
Immunogen
This IgY fraction antibody was prepared from eggs of chickens laid after repeated immunizations with a synthetic peptide corresponding to aa 2-32 of Human MIF conjugated to keyhole limpet hemocyanin (KLH). MIF is a proinflammatory cytokine that plays an important role in systemic inflammatory events.