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Key Product Details

Species Reactivity

Validated:

Rat

Cited:

Rat

Applications

Validated:

CyTOF-ready, Immunocytochemistry, Intracellular Staining by Flow Cytometry, Neutralization, Western Blot

Cited:

Flow Cytometry, Immunohistochemistry-Paraffin

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 83308

Product Specifications

Immunogen

E. coli-derived recombinant rat GM-CSF
Ala1-Lys127
Accession # P48750

Specificity

Detects rat GM-CSF in direct ELISAs and Western blots. In direct ELISAs and Western blots, no cross-reactivity with recombinant GM‑CSF from mouse, human, or pig is observed.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Rat GM-CSF Antibody

Cell Proliferation Induced by GM‑CSF and Neutralization by Rat GM‑CSF Antibody.

Cell Proliferation Induced by GM‑CSF and Neutralization by Rat GM‑CSF Antibody.

Recombinant Rat GM-CSF (Catalog # 518-GM) stimulates proliferation in the DA3 mouse myeloma cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Rat GM-CSF (0.5 ng/mL) is neutralized (green line) by increasing concentrations of Rat GM-CSF Monoclonal Antibody (Catalog # MAB5181). The ND50 is typically 1-4 µg/mL.
GM-CSF antibody in Rat Splenocytes by Immunocytochemistry (ICC).

GM‑CSF in Rat Splenocytes.

GM-CSF was detected in immersion fixed rat splenocytes using Rat GM-CSF Monoclonal Antibody (Catalog # MAB5181) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Applications for Rat GM-CSF Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed rat splenocytes

Intracellular Staining by Flow Cytometry

2.5 µg/106 cells
Sample: Rat splenocytes fixed with paraformaldehyde and permeabilized with saponin

Western Blot

1 µg/mL
Sample: Recombinant Rat GM-CSF (Catalog # 518-GM) under non-reducing conditions only. Catalog # AF518 is recommended to detect rat GM-CSF in Western blots.

Neutralization

Measured by its ability to neutralize GM‑CSF-induced proliferation in the DA3 mouse myeloma cell line. Ihle, J. N. et al. (1984) Advances in Viral Oncology. In G. Klein (eds): Raven Press, New York, NY. 4:95. The Neutralization Dose (ND50) is typically 1-4 µg/mL in the presence of 0.5 ng/mL Recombinant Rat GM‑CSF.

Reviewed Applications

Read 1 review rated 5 using MAB5181 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Reconstitution Buffer Available:
Size / Price
Qty
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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 rat GM-CSF shares 56%‑69% amino acid sequence identity with canine, feline, human, mouse, 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). Rat GM‑CSF is active on mouse cells, although mouse GM‑CSF is only weakly active on rat 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. Smith, L.R. et al. (1994) Immunogenetics 39:80.
  11. Onetto-Pothier, N. et al. (1990) Blood 75:59.
  12. Hayashida, K. et al. (1990) Proc. Natl. Acad. Sci. 87:9655.
  13. Pelley, J.L. et al. (2007) Exp. Hematol. 35:1483.
  14. Oaks, M.K. et al. (1995) J. Interferon Cytokine Res. 15:1095.
  15. Vandenabeele, P. et al. (1990) Lymphokine Res. 9:381.

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 Rat 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 Rat GM-CSF Antibody

For research use only

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