Canine GM-CSF Antibody [Biotin]

R&D Systems | Catalog # BAF1546

R&D Systems

Key Product Details

Species Reactivity

Canine

Applications

Western Blot, ELISA Detection (Matched Antibody Pair), Immunocytochemistry

Label

Biotin

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 ELISAs and Western blots. In sandwich immunoassays, less than 0.2% cross-reactivity with recombinant human GM-CSF, recombinant mouse GM-CSF, recombinant feline GM-CSF, and recombinant canine IL-5 is observed and less than

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Canine GM-CSF Antibody [Biotin]

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) stimulated with calcium ionomycin and PMA using Goat Anti-Canine GM-CSF Biotinylated Antigen Affinity-purified Polyclonal Antibody (Catalog # BAF1546) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Streptavidin (red; Catalog # NL999) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Canine GM-CSF ELISA Standard Curve

Canine GM-CSF ELISA Standard Curve

Recombinant Canine GM‑CSF (Catalog # 1546-GM) was serially diluted and captured by Mouse Anti-Canine GM‑CSF Monoclonal Antibody (Catalog # MAB1546) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Canine GM‑CSF Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1546) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)

Applications for Canine GM-CSF Antibody [Biotin]

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed canine peripheral blood mononuclear cells (PBMCs) stimulated with calcium ionomycin and PMA

Western Blot

0.1 µg/mL
Sample: Recombinant Canine GM-CSF (Catalog # 1546-GM)

Canine GM-CSF Sandwich Immunoassay

ELISA Detection (Matched Antibody Pair)
Recommended Concentration: 0.1-0.4 µg/mL
Use in combination with these reagents:
  • Capture Reagent: Canine GM‑CSF Antibody (Catalog # MAB1546)
  • Standard: Recombinant Canine GM-CSF Protein (Catalog # 1546-GM)
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.


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Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.

Shipping

The product is shipped at ambient temperature. Upon receipt, store it 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 [Biotin]

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Canine GM-CSF Antibody [Biotin]

For research use only

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Protocols

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