Canine GM-CSF Antibody

(2 citations)   
  • Species Reactivity
    Canine
  • Specificity
    Detects canine GM-CSF in direct ELISAs and Western blots. In Western blots, approximately 25% cross-reactivity with recombinant rat GM‑CSF is observed, approximately 5% cross-reactivity with recombinant feline GM-CSF and recombinant porcine GM‑CSF is observed, and less than 1% cross-reactivity with recombinant human GM‑CSF and recombinant mouse GM‑CSF is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant canine GM-CSF
    Ala18-Lys144
    Accession # P48749.1
  • Formulation
    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.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.1 µg/mL
    Recombinant Canine GM-CSF (Catalog # 1546-GM)
  • Immunocytochemistry
    5-15 µg/mL
    See below
  • Neutralization
    Measured by its ability to neutralize GM‑CSF-induced proliferation in the TF‑1 human erythroleukemic cell line. Kitamura, T. et al. (1989) J. Cell Physiol. 140:323. The Neutralization Dose (ND50) is typically 2-8 µg/mL in the presence of 15 ng/mL Recombinant Canine 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.
Data Examples
Cell Proliferation Induced by GM‑CSF and Neutralization by Canine GM‑CSF Antibody.

Recombinant Canine GM‑CSF (Catalog # 1546-GM) stimulates proliferation in the TF‑1 human erythroleukemic cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Canine GM‑CSF (15 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Canine GM‑CSF Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1546). The ND50 is typically 2-8 µg/mL.

Immunocytochemistry
GM‑CSF in Canine PBMCs. GM‑CSF was detected in immersion fixed canine peripheral blood mononuclear cells (PBMCs) treated with Calcium Ionomycin and PMA using Goat Anti-Canine GM‑CSF Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1546) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) 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
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    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.
Background: GM-CSF

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 canine GM-CSF shares 49-57% amino acid sequence identity with mouse and rat GM-CSF and 69-72% with feline, human, 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). The activity of GM‑CSF is species specific between human and mouse, although human GM‑CSF is active on canine cells (14, 15).

  • References:
    1. Martinez-Moczygemba, M. and D.P. Huston (2003) J. Allergy Clin. Immunol. 112:653.
    2. Barreda, D.R. et al. (2004) Dev. Comp. Immunol. 28:509. 
    3. Eksioglu, E.A. et al. (2007) Exp. Hematol. 35:1163. 
    4. Cao, Y. (2007) J. Clin. Invest. 117:2362.
    5. Fleetwood, A.J. et al. (2005) Crit. Rev. Immunol. 25:405.
    6. Heuser, M. et al. (2007) Semin. Hematol. 44:148.
    7. Hege, K.M. et al. (2006) Int. Rev. Immunol. 25:321.
    8. Kaushansky, K. et al. (1992) Biochemistry 31:1881.
    9. Diederichs, K. et al. (1991) Science 254:1779.
    10. Nash, R.A. et al. (1991) Blood 78:930.
    11. Onetto-Pothier, N. et al. (1990) Blood 75:59.
    12. Hayashida, K. et al. (1990) Proc. Natl. Acad. Sci. USA 87:9655.
    13. Pelley, J.L. et al. (2007) Exp. Hematol. 35:1483.
    14. Shanafelt, A.B. et al. (1991) J. Biol. Chem. 266:13804.
    15. Hogge, G.S. et al. (1990) Cancer Gene Ther. 6:26.
  • Long Name:
    Granulocyte Macrophage Growth Factor
  • Entrez Gene IDs:
    1437 (Human); 12981 (Mouse); 116630 (Rat); 397208 (Porcine); 403923 (Canine); 493805 (Feline)
  • Alternate Names:
    colony stimulating factor 2 (granulocyte-macrophage); Colony-stimulating factor; CSF; CSF2; GMCSF; GM-CSF; GMCSFgranulocyte-macrophage colony-stimulating factor; granulocyte-macrophage colony stimulating factor; MGC131935; MGC138897; molgramostin; sargramostim
Related Research Areas
Citations:

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.

2 Citations: Showing 1 - 2
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Species
Applications
Sample Type
  1. Preventive and therapeutic effects of gene therapy using silica nanoparticles-binding of GM-CSF gene on white blood cell production in dogs with leukopenia.
    Authors: Choi EW, Koo HC, Shin IS, Chae YJ, Lee JH, Han SM, Lee SJ, Bhang DH, Park YH, Lee CW, Youn HY
    Exp. Hematol., 2008;36(9):1091-7.
    Species: Canine
    Sample Type: Serum
    Application: ELISA Development
  2. Effects of GM-CSF gene transfer using silica-nanoparticles as a vehicle on white blood cell production in dogs.
    Authors: Choi EW, Shin IS, Chae YJ, Koo HC, Lee JH, Chung TH, Park YH, Kim DY, Hwang CY, Lee CW, Youn HY
    Exp. Hematol., 2008;36(7):807-15.
    Species: Canine
    Sample Type: Serum
    Application: ELISA Development
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