Detects rat GM-CSF in direct ELISAs and Western blots. In direct ELISAs and Western blots, no cross-reactivity with recombinant GM‑CSF from mouse, human, or pig is observed.
Monoclonal Mouse IgG2B Clone # 83308
Protein A or G purified from hybridoma culture supernatant
E. coli-derived recombinant rat GM-CSF Ala1-Lys127 Accession # P48750
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
<0.10 EU per 1 μg of the antibody by the LAL method.
Recombinant Rat GM-CSF (Catalog # 518-GM) under non-reducing conditions only. Catalog # AF518 is recommended to detect rat GM-CSF in Western blots.
Intracellular Staining by Flow Cytometry
2.5 µg/106 cells
Rat splenocytes fixed with paraformaldehyde and permeabilized with saponin
Measured by its ability to neutralize GM‑CSF-induced proliferation in the DA3 mouse myeloma cell line. Ihle, J. N. et al. (1984) Advances in Viral Oncology. In G. Klein (eds): Raven Press, New York, NY. 4:95. The Neutralization Dose (ND50) is typically 1-4 µg/mL in the presence of 0.5 ng/mL Recombinant Rat GM‑CSF.
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.
Cell Proliferation Induced by GM‑CSF and Neutralization by Rat GM‑CSF Antibody. Recombinant Rat GM‑CSF (Catalog # 518-GM) stimulates proliferation in the DA3 mouse myeloma cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Rat GM‑CSF (0.5 ng/mL) is neutralized (green line) by increasing concentrations of Rat GM‑CSF Monoclonal Antibody (Catalog # MAB5181). The ND50 is typically 1-4 µg/mL.
GM‑CSF in Rat Splenocytes. GM‑CSF was detected in immersion fixed rat splenocytes using Rat GM‑CSF Monoclonal Antibody (Catalog # MAB5181) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Preparation and Storage
Reconstitute at 0.5 mg/mL in sterile PBS.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
GM-CSF was initially characterized as a factor that can support the in vitro colony formation of granulocyte-macrophage progenitors. It is also a growth factor for erythroid, megakaryocyte, and eosinophil progenitors. GM-CSF is produced by a number of different cell types (including T cells, B cells, macrophages, mast cells, endothelial cells, fibroblasts, and adipocytes) in response to cytokine or inflammatory stimuli. On mature hematopoietic cells, GM-CSF is a survival factor for and activates the effector functions of granulocytes, monocytes/macrophages, and eosinophils (1, 2). GM-CSF promotes a Th1 biased immune response, angiogenesis, allergic inflammation, and the development of autoimmunity (3‑5). It shows clinical effectiveness in ameliorating chemotherapy-induced neutropenia, and GM-CSF transfected tumor cells are utilized as cancer vaccines (6, 7). The 22 kDa glycosylated GM-CSF, similar to IL‑3 and IL‑5, is a cytokine with a core of four bundled alpha ‑helices (8‑10). Mature rat GM-CSF shares 56%‑69% amino acid sequence identity with canine, feline, human, mouse, and porcine GM‑CSF. GM‑CSF exerts its biological effects through a heterodimeric receptor complex composed of GM‑CSF R alpha /CD116 and the signal transducing common beta chain (CD131) which is also a component of the high-affinity receptors for IL-3 and IL-5 (11, 12). In addition, GM-CSF binds a naturally occurring soluble form of GM‑CSF R alpha (13). Rat GM‑CSF is active on mouse cells, although mouse GM‑CSF is only weakly active on rat cells (14, 15).
Martinez-Moczygemba, M. and D.P. Huston (2003) J. Allergy Clin. Immunol. 112:653.
Barreda, D.R. et al. (2004) Dev. Comp. Immunol. 28:509.
Eksioglu, E.A. et al. (2007) Exp. Hematol. 35:1163.
Cao, Y. (2007) J. Clin. Invest. 117:2362.
Fleetwood, A.J. et al. (2005) Crit. Rev. Immunol. 25:405.
Heuser, M. et al. (2007) Semin. Hematol. 44:148.
Hege, K.M. et al. (2006) Int. Rev. Immunol. 25:321.
Kaushansky, K. et al. (1992) Biochemistry 31:1881.
Diederichs, K. et al. (1991) Science 254:1779.
Smith, L.R. et al. (1994) Immunogenetics 39:80.
Onetto-Pothier, N. et al. (1990) Blood 75:59.
Hayashida, K. et al. (1990) Proc. Natl. Acad. Sci. 87:9655.
Pelley, J.L. et al. (2007) Exp. Hematol. 35:1483.
Oaks, M.K. et al. (1995) J. Interferon Cytokine Res. 15:1095.
Vandenabeele, P. et al. (1990) Lymphokine Res. 9:381.
R&D Systems personnel manually curate a database that contains references using R&D Systems products.
The data collected includes not only links to publications in PubMed,
but also provides information about sample types, species, and experimental conditions.
The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.