GM-CSF Antibody [Unconjugated]

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 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.

• Reactivity: Ca
• Applications: WB, B/N, ICC/IF
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

GM-CSF Antibody [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant canine GM-CSF
  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.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: CSF2
• Purity Statement: Antigen Affinity-purified
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 5-15 ug/mL
  • Neutralization 2-8 ug/mL
  • Western Blot 0.1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for GM-CSF Antibody [Unconjugated]

• colony stimulating factor 2 (granulocyte-macrophage)
• Colony-stimulating factor
• CSF
• CSF2
• CSF-2
• GMCSF
• GM-CSF
• GMCSFgranulocyte-macrophage colony-stimulating factor
• granulocyte-macrophage colony stimulating factor
• MGC131935
• MGC138897
• Molgramostim
• molgramostin
• Sargramostim

Background

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).

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for GM-CSF Antibody (AF1546)(4)

Publications using AF1546

• K Marek, F Armando, VM Nippold, K Rohn, P Plattet, G Brogden, G Gerold, W Baumgärtne, C Puff Persistent Infection of a Canine Histiocytic Sarcoma Cell Line with Attenuated Canine Distemper Virus Expressing Vasostatin or Granulocyte-Macrophage Colony-Stimulating Factor International Journal of Molecular Sciences, 2022-05-31;23(11):. 2022-05-31 [PMID: 35682834] (Western Blot, Canine)
• Wang C, Ke H et al. Protective effect of nicorandil on collapse induced lung injury in rabbits by inhibiting apoptosis. Int J Mol Med 1/8/2019 [PMID: 31173181]
• Choi EW, Koo HC, Shin IS, Chae YJ, Lee JH, Han SM, Lee SJ, Bhang DH, Park YH, Lee CW, Youn HY Preventive and therapeutic effects of gene therapy using silica nanoparticles-binding of GM-CSF gene on white blood cell production in dogs with leukopenia. Exp. Hematol., 2008-06-11;36(9):1091-7. 2008-06-11 [PMID: 18550260] (ELISA Development, Canine)
• Choi EW, Shin IS, Chae YJ, Koo HC, Lee JH, Chung TH, Park YH, Kim DY, Hwang CY, Lee CW, Youn HY Effects of GM-CSF gene transfer using silica-nanoparticles as a vehicle on white blood cell production in dogs. Exp. Hematol., 2008-04-02;36(7):807-15. 2008-04-02 [PMID: 18375041] (ELISA Development, Canine)

Bioinformatics

• Gene Symbol: CSF2
• Uniprot:
  • P48749.1()