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

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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

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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

Citations for Canine GM‑CSF Antibody

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Protocols

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