Detection of Fc gamma RI/CD64 in RAW 264.7 Mouse Cell Line by Flow Cytometry.
RAW 264.7 mouse monocyte/macrophage cell line was stained with Rat Anti-Mouse Fc gamma RI/CD64 PerCP‑conjugated Monoclonal Antibody (Catalog # FAB20741C, filled histogram) or isotype control antibody (Catalog # IC006C, open histogram). View our protocol for Staining Membrane-associated Proteins.
Preparation and Storage
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Protect from light. Do not freeze.
12 months from date of receipt, 2 to 8 °C as supplied.
Background: Fc gamma RI/CD64
Receptors for the Fc region of IgG (Fc gamma Rs) are members of the Ig superfamily that function in the activation or inhibition of immune responses such as degranulation, phagocytosis, ADCC (antibody-dependent cellular toxicity), cytokine release, and B cell proliferation (1‑3). The Fc gamma Rs have been divided into three classes based on close relationships in their extracellular domains; these groups are designated Fc gamma RI (also known as CD64), Fc gamma RII (CD32), and Fc gamma RIII (CD16). Each group may be encoded by multiple genes and exist in different isoforms depending on species and cell type. The CD64 proteins are high affinity receptors (~10‑8‑10‑9 M) capable of binding monomeric IgG, whereas the CD16 and CD32 proteins bind IgG with lower affinities (~10‑6‑10‑7 M) only recognizing IgG aggregates surrounding multivalent antigens (1, 4). Fc gamma Rs that deliver an activating signal either have an intrinsic immunoreceptor tyrosine-based activation motif (ITAM) within their cytoplasmic domains or associate with one of the ITAM-bearing adapter subunits, Fc R gamma or zeta (3, 5). The only inhibitory member in human and mouse, Fc gamma RIIb, has an intrinsic cytoplasmic immunoreceptor tyrosine-based inhibitory motif (ITIM). The coordinated functioning of activating and inhibitory receptors is necessary for successful initiation, amplification, and termination of immune responses (5). Mouse Fc gamma RI is transmembrane protein with three extracellular Ig-like domains, and it delivers an activating signal via the associated Fc R gamma accessory chain (1, 2). The high affinity recognition of IgG by Fc gamma RI permits the triggering of effector responses at low IgG concentrations typical of early immune responses (2). Fc gamma RI is expressed constitutively on monocytes and macrophages and can be induced on neutrophils and eosinophils (1, 4). Its expression is up-regulated during bacterial infections and sepsis.
Van de Winkel, J. and P. Capes (1993) Immunol. Today 14:215.
Raghaven, M. and P. Bjorkman (1996) Annu. Rev. Cell Dev. Biol. 12:181.
Ravetch, J. and S. Bolland (2001) Annu. Rev. Immunol. 19:275.
CD64 antigen; CD64; Fc fragment of IgG, high affinity Ia, receptor (CD64); Fc gamma RI; FCG1; Fc-gamma receptor I B2; Fc-gamma RI; Fc-gamma RIA; FcgammaRIa; FCGR1; FcgRI; FCRI; FLJ18345; high affinity Ia, receptor for (CD64); high affinity immunoglobulin gamma Fc receptor I; IgG Fc receptor I
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Other Experimental Details
High content methanol (80%) based protocol. In two different experiments the separation of a positive and negative population was not as clear as I would have needed for my research question.