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Canine GM-CSF Antibody

R&D Systems, part of Bio-Techne | Catalog # AF1546

R&D Systems, part of Bio-Techne
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AF1546
AF1546-SP

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Canine

Cited:

Canine

Applications

Validated:

Immunocytochemistry, Neutralization, Western Blot

Cited:

ELISA Development, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

Product Specifications

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.

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.

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 antibody in Canine PBMCs by Immunocytochemistry (ICC).

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

Western Blot

0.1 µg/mL
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.

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

Alternate Names

CSF-2, CSF2, GMCSF, Molgramostim, Sargramostim

Entrez Gene IDs

1437 (Human); 12981 (Mouse); 116630 (Rat); 397208 (Porcine); 403923 (Canine); 493805 (Feline)

Gene Symbol

CSF2

UniProt

Additional GM-CSF Products

Product Documents for Canine GM-CSF Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Canine GM-CSF Antibody

For research use only

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