Canine GM-CSF Biotinylated Antibody

Catalog #: BAF1546 Datasheet

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BAF1546

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Canine GM-CSF Biotinylated Antibody Summary

Species Reactivity

Canine

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
0.7% cross‑reactivity with recombinant porcine GM-CSF is observed.

Source

Polyclonal Goat IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant canine GM-CSF
Ala18-Lys144
Accession # P48749.1

Formulation

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

Label

Biotin

Applications

Recommended Concentration

Sample

Western Blot

0.1 µg/mL

Recombinant Canine GM-CSF (Catalog # 1546-GM)

Immunocytochemistry

5-15 µg/mL

See below

Canine GM-CSF Sandwich Immunoassay

Recommended Concentration

Reagent

ELISA Detection (Matched Antibody Pair)

0.1-0.4 µg/mL

Use in combination with:

Capture Reagent: Canine GM-CSF Antibody (Catalog # MAB1546)

Standard: Recombinant Canine GM-CSF Protein (Catalog # 1546-GM)

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Immunocytochemistry

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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.

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

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.

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

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.
Nash, R.A. et al. (1991) Blood 78:930.
Onetto-Pothier, N. et al. (1990) Blood 75:59.
Hayashida, K. et al. (1990) Proc. Natl. Acad. Sci. USA 87:9655.
Pelley, J.L. et al. (2007) Exp. Hematol. 35:1483.
Shanafelt, A.B. et al. (1991) J. Biol. Chem. 266:13804.
Hogge, G.S. et al. (1990) Cancer Gene Ther. 6:26.

Long Name

Granulocyte Macrophage Growth Factor

Entrez Gene IDs

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

Alternate Names

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