Key Product Details

Species Reactivity

Validated:

Rat

Cited:

Rat

Applications

Validated:

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

Cited:

Immunohistochemistry-Paraffin, Flow Cytometry

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 83308
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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:

Flow Cytometry Panel Builder

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Save time and reduce costly mistakes by quickly finding compatible reagents using the Panel Builder Tool.

Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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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.


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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 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 or batch 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

Citations for Rat GM‑CSF Antibody

Customer Reviews for Rat GM‑CSF Antibody (1)

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  • Rat GM-CSF Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: Splenocytes
    Species: Rat
    Verified Customer | Posted 03/15/2022
    Rat GM‑CSF Antibody MAB5181

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Protocols

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