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).
Canine GM‑CSF Antibody
R&D Systems | Catalog # AF1546
Key Product Details
Validated by
Biological Validation
Species Reactivity
Validated:
Canine
Cited:
Canine
Applications
Validated:
Western Blot, Neutralization, Immunocytochemistry
Cited:
Western Blot, ELISA Development
Label
Unconjugated
Antibody Source
Polyclonal Goat IgG
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Product Specifications
Immunogen
E. coli-derived recombinant canine GM-CSF
Ala18-Lys144
Accession # P48749.1
Ala18-Lys144
Accession # P48749.1
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.
Clonality
Polyclonal
Host
Goat
Isotype
IgG
Endotoxin Level
<0.10 EU per 1 μg of the antibody by the LAL method.
Scientific Data Images for Canine GM‑CSF Antibody
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.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.Applications for Canine GM‑CSF Antibody
Application
Recommended Usage
Immunocytochemistry
5-15 µg/mL
Sample: Immersion fixed canine peripheral blood mononuclear cells (PBMCs) treated with calcium ionomycin and PMA
Sample: Immersion fixed canine peripheral blood mononuclear cells (PBMCs) treated with calcium ionomycin and PMA
Western Blot
0.1 µg/mL
Sample: Recombinant Canine GM-CSF (Catalog # 1546-GM)
Sample: Recombinant Canine GM-CSF (Catalog # 1546-GM)
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.
Formulation, Preparation, and Storage
Purification
Antigen Affinity-purified
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
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Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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.
Calculators
Background: GM-CSF
References
- 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.
- Nash, R.A. et al. (1991) Blood 78:930.
- Onetto-Pothier, N. et al. (1990) Blood 75:59.
- Hayashida, K. et al. (1990) Proc. Natl. Acad. Sci. USA 87:9655.
- Pelley, J.L. et al. (2007) Exp. Hematol. 35:1483.
- Shanafelt, A.B. et al. (1991) J. Biol. Chem. 266:13804.
- Hogge, G.S. et al. (1990) Cancer Gene Ther. 6:26.
Long Name
Granulocyte Macrophage Growth Factor
Alternate Names
CSF-2, CSF2, GMCSF, Molgramostim, Sargramostim
Entrez Gene IDs
Gene Symbol
CSF2
UniProt
Additional GM-CSF Products
Product Documents for Canine GM‑CSF Antibody
Certificate of Analysis
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Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Canine GM‑CSF Antibody
For research use only
Related Research Areas
Citations for Canine GM‑CSF Antibody
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- ClariTSA™ Fluorophore Kits
- Detection & Visualization of Antibody Binding
- ICC Cell Smear Protocol for Suspension Cells
- ICC Immunocytochemistry Protocol Videos
- ICC for Adherent Cells
- Immunocytochemistry (ICC) Protocol
- Immunocytochemistry Troubleshooting
- Immunofluorescence of Organoids Embedded in Cultrex Basement Membrane Extract
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Fluorescent ICC Staining of Cell Smears - Graphic
- Protocol for the Fluorescent ICC Staining of Cultured Cells on Coverslips - Graphic
- Protocol for the Preparation and Fluorescent ICC Staining of Cells on Coverslips
- Protocol for the Preparation and Fluorescent ICC Staining of Non-adherent Cells
- Protocol for the Preparation and Fluorescent ICC Staining of Stem Cells on Coverslips
- Protocol for the Preparation of a Cell Smear for Non-adherent Cell ICC - Graphic
- R&D Systems Quality Control Western Blot Protocol
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars