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Ionized calcium binding adaptor molecule 1 (IBA1) is a 147 amino acid macrophage- and microglial-specific calcium binding protein, widely used as a marker for microglia. This preparation of rabbit polyclonal IgG antibody is qualified for immunocytochemical staining of human microglia and supplied at 0.5mg/ml in tris-buffered saline (TBS). The polyclonal antibody is cross-reactive to Iba1 from a wide range of species including human, mouse, rat, etc. The antibody is supplied unconjugated, permitting your choice of secondary detection methods.
Immunofluorescent labeling is a straight-forward technique to assessing the presence and subcellular localization of an antigen or a protein. Several labeling methods are available depending on the biological sample, cell preparation, and availability of antibodies against the target. The method presented in our protocol has demonstrated utility in detecting the presence of triggering receptor expressed on myeloid cells 2 (TREM2), ionized calcium-binding adapter molecule 1 (IBA1), CX3-C motif chemokine receptor 1 (CX3CR1), spi-1 proto-oncogene (PU.1), purinergic receptor P2Y12 (P2RY12), and integrin subunit alpha M (CD11b) in iCell® Microglia. The provided protocol describes plating and labeling of iCell Microglia for fluorescent immunocytochemistry assays.
Plated, fixed microglia were immunofluorescently labeled using anti IBA1 (green) and Trem2 (red). Cells were plated at approximately 14,500 cells/well in a 384 well plate.