Mouse GM-CSF R alpha PE-conjugated Antibody Summary
Accession # Q00941
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Detection of GM-CSF R alpha in J774A.1 Mouse Cell Line by Flow Cytometry. J774A.1 mouse reticulum cell sarcoma macrophage cell line was stained with Rat Anti-Mouse GM-CSF Ra PE-conjugated Monoclonal Antibody (Catalog # FAB6130P, filled histogram) or isotype control antibody (Catalog # IC006P, open histogram). View our protocol for Staining Membrane-associated Proteins.
Preparation and Storage
Background: GM-CSF R alpha
Granulocyte macrophage colony stimulating factor receptor alpha (GM-CSF R alpha ), also known as CD116, is a component of the receptor complex that mediates cellular responses to GM-CSF. GM-CSF promotes the differentiation and mobilization of granulocyte-macrophage, erythroid, megakaryocyte, and eosinophil progenitors. It enhances the activation of myeloid cell effector functions and plays a role in the development of Th1 biased immune responses, allergic inflammation, and autoimmunity (1-4). Mature mouse GM-CSF R alpha is an 80 kDa type I transmembrane glycoprotein that consists of a 298 amino acid (aa) extracellular domain (ECD) with two fibronectin type III domains and a juxtamembrane WSXWS motif, a 21 aa transmembrane segment, and a 40 aa cytoplasmic domain (5). Within the ECD, mouse GM-CSF R alpha shares approximately 33% and 58% aa sequence identity with human and rat GM-CSF R alpha, respectively. Soluble forms of the human receptor retain the ability to bind GM-CSF (6, 7). GM-CSF R alpha is expressed on hematopoietic stem cells, progenitor and differentiated cells in the myeloid lineage, vascular endothelial cells, placenta, and non‑hematopoietic solid tumor cells (8). GM-CSF R alpha associates with the common beta chain/CD131 ( beta c), a 135 kDa transmembrane protein that is also the signal transducing component of the receptors for IL-3 and IL-5 (9, 10). Association with beta c converts GM-CSF R alpha from a low affinity to a high affinity receptor for GM-CSF (9-11). The shared usage of beta c underlies the synergism between GM-CSF, IL-3, and IL-5 in their effects on myeloid cell differentiation and activation (1, 2).
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- Hoang, T. et al. (1993) J. Biol. Chem. 268:11881.
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Neutrophils were differentiated in vitro from granulocyte/macrophage progenitors. Cells were stained for neutrophil surface markers (CD11b, Gr-1) and populations show gated neutrophils. Anti-GM-CSFR alpha antibody was titrated in the surface stain or after surface staining with intracellular staining after cells were fixed and permeablized. Like for many surface receptors, the antibody did not stain well when use to stain the surface of cells; however, a positive stain was seen with intracellular staining. There isn't a huge difference between positive staining and the FMO control minimizing the applications for this antibody for flow cytometry.