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).
Key Product Details
Species Reactivity
Validated:
Rat
Cited:
Rat
Applications
Validated:
Western Blot, Neutralization, Intracellular Staining by Flow Cytometry, Immunocytochemistry, CyTOF-ready
Cited:
Immunohistochemistry-Paraffin, Flow Cytometry
Label
Unconjugated
Antibody Source
Monoclonal Mouse IgG2B Clone # 83308
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Product Specifications
Immunogen
E. coli-derived recombinant rat GM-CSF
Ala1-Lys127
Accession # P48750
Ala1-Lys127
Accession # P48750
Specificity
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.
Clonality
Monoclonal
Host
Mouse
Isotype
IgG2B
Endotoxin Level
<0.10 EU per 1 μg of the antibody by the LAL method.
Scientific Data Images for Rat GM‑CSF Antibody
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.Applications for Rat GM‑CSF Antibody
Application
Recommended Usage
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
Immunocytochemistry
8-25 µg/mL
Sample: Immersion fixed rat splenocytes
Sample: Immersion fixed rat splenocytes
Intracellular Staining by Flow Cytometry
2.5 µg/106 cells
Sample: Rat splenocytes fixed with paraformaldehyde and permeabilized with saponin
Sample: Rat splenocytes fixed with paraformaldehyde and permeabilized with saponin
Western Blot
1 µg/mL
Sample: Recombinant Rat GM-CSF (Catalog # 518-GM) under non-reducing conditions only. Catalog # AF518 is recommended to detect rat GM-CSF in Western blots.
Sample: Recombinant Rat GM-CSF (Catalog # 518-GM) under non-reducing conditions only. Catalog # AF518 is recommended to detect rat GM-CSF in Western blots.
Neutralization
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.
Reviewed Applications
Read 1 review rated 5 using MAB5181 in the following applications:
Flow Cytometry Panel Builder
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Advanced Features
- Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
- Spillover Popups - Visualize the spectra of individual fluorochromes
- Antigen Density Selector - Match fluorochrome brightness with antigen density
Formulation, Preparation, and Storage
Purification
Protein A or G purified from hybridoma culture supernatant
Reconstitution
Reconstitute at 0.5 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.
- 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.
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 Rat GM‑CSF Antibody
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Rat GM‑CSF Antibody
For research use only
Related Research Areas
Citations for Rat GM‑CSF Antibody
Customer Reviews for Rat GM‑CSF Antibody (1)
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Application: Immunocytochemistry/ImmunofluorescenceSample Tested: SplenocytesSpecies: RatVerified Customer | Posted 03/15/2022
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- 7-Amino Actinomycin D (7-AAD) Cell Viability Flow Cytometry Protocol
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- ClariTSA™ Fluorophore Kits
- Detection & Visualization of Antibody Binding
- Extracellular Membrane Flow Cytometry Protocol
- Flow Cytometry Protocol for Cell Surface Markers
- Flow Cytometry Protocol for Staining Membrane Associated Proteins
- Flow Cytometry Staining Protocols
- Flow Cytometry Troubleshooting Guide
- 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
- Intracellular Flow Cytometry Protocol Using Alcohol (Methanol)
- Intracellular Flow Cytometry Protocol Using Detergents
- Intracellular Nuclear Staining Flow Cytometry Protocol Using Detergents
- Intracellular Staining Flow Cytometry Protocol Using Alcohol Permeabilization
- Intracellular Staining Flow Cytometry Protocol Using Detergents to Permeabilize Cells
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Propidium Iodide Cell Viability Flow Cytometry Protocol
- Protocol for Liperfluo
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Characterization of Human Th22 Cells
- Protocol for the Characterization of Human Th9 Cells
- 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
- Protocol: Annexin V and PI Staining by Flow Cytometry
- Protocol: Annexin V and PI Staining for Apoptosis by Flow Cytometry
- 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: Fluorokine Flow Cytometry Kits
- 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