Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

Western Blot, Neutralization, Immunocytochemistry

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Neutralization, Immunodepletion

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 3209
View all GM-CSF Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human GM-CSF
Ala18-Glu144
Accession # P04141

Specificity

Detects human GM-CSF in Western blots.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Endotoxin Level

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

Scientific Data Images for Human GM‑CSF Antibody

GMCSF antibody in Human PBMCs by Immunocytochemistry (ICC).

GMCSF in Human PBMCs.

Granulocyte Macrophage Colony Stimulating Factor (GM-CSF) was detected in human peripheral blood mononuclear cells (PBMCs) using Human GM-CSF Monoclonal Antibody (Catalog # MAB215) at 10 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). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Cell Proliferation Induced by GM‑CSF and Neutralization by Human GM‑CSF Antibody.

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

Recombinant Human GM-CSF (Catalog # 215-GM) stimulates proliferation in the TF-1 human erythroleukemic cell line in a dose-dependent manner (orange line) as measured by Resazurin (Catalog # AR002). Proliferation elicited by Recombinant Human GM-CSF (0.1 ng/mL) is neutralized (green line) by increasing concentrations of Human GM-CSF Monoclonal Antibody (Catalog # MAB215). The ND50 is typically 0.075-0.45 µg/mL.
Detection of GM-CSF by Flow Cytometry

Detection of GM-CSF by Flow Cytometry

TC-derived soluble factors promoted neutrophil survival. A. Neutrophils were cultured in a TC-CM or the control medium. At the indicated time points, live cells were evaluated by flow cytometry with FITC-conjugated annexin V and PI. Results were expressed as percentages of live cells (mean ± SEM of five independent experiments); ***p < 0.005; **p < 0.01; *p < 0.05. B. Representative flow cytometric panels of dot plots of PMNs cultured in a TC-CM or control medium and stained with FITC-conjugated annexin V and propidium iodide (PI) at 24 (upper panels) and 48 (lower panels) hours. C. The GM-CSF release by TPC1 and 8505c cells was evaluated by an ELISA in a TC-CM or in the control medium. Results were expressed as mean ± SEM of seven independent experiments; ****p < 0.001; ***p < 0.005. D-F. Neutrophil survival in a TPC1-derived (D-E) or 8505c-derived (F-G) conditioned medium was evaluated in the presence of an anti-GM-CSF blocking antibody or the relative isotype control (10 μg/ml). At 24 hours, live cells were stained with FITC-conjugated annexin V and PI and analyzed by flow cytometry. Figs E and G illustrate representative flow cytometric panels of one out of five independent experiments. The results were expressed as mean ± SEM of five independent experiments; **p < 0.01; *p < 0.05. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/29953504), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of GM-CSF by Immunohistochemistry

Detection of GM-CSF by Immunohistochemistry

Knockdown of GM-CSF protein levels after siRNA application in cancer cells. HeLa/DLD-1 cells were transfected with control siRNA (1/1*, 2/2*) or GM-CSF siRNA (3/3*, 4/4*) and cultured in the absence or presence of 25 ng/mL HB-EGF. The numbers indicate the culture conditions and the corresponding supernatants (SN) used for ELISA or cell stimulation. (A) Blockade of GM-CSF production in cultures of HeLa/DLD-1 cells transfected with GM-CSF siRNA was confirmed by immunocytochemistry (2/2* vs. 4/4*) and ELISA (left side; 2/2* vs. 4/4*, p < 0.05). (B) SN from GM-CSF-silenced HeLa/DLD-1 did not induce HB-EGF expression in mononuclear phagocytes (Mø), as revealed by flow cytometry (2/2* vs. 4/4*) and ELISA (left side; 2/2* vs. 4/4*, p < 0.05). (C) Mø stimulated with SN from GM-CSF-silenced HeLa/DLD-1 cells released SN less effective at inducing GM-CSF in non-silenced cancer cells, as determined by ELISA (see Methods section; SN2 vs. SN4, p < 0.05). Representative pictures or the means ± SD out of 5 experiments are shown. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/20946648), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of GM-CSF by Flow Cytometry

Detection of GM-CSF by Flow Cytometry

TC-derived soluble factors promoted neutrophil survival. A. Neutrophils were cultured in a TC-CM or the control medium. At the indicated time points, live cells were evaluated by flow cytometry with FITC-conjugated annexin V and PI. Results were expressed as percentages of live cells (mean ± SEM of five independent experiments); ***p < 0.005; **p < 0.01; *p < 0.05. B. Representative flow cytometric panels of dot plots of PMNs cultured in a TC-CM or control medium and stained with FITC-conjugated annexin V and propidium iodide (PI) at 24 (upper panels) and 48 (lower panels) hours. C. The GM-CSF release by TPC1 and 8505c cells was evaluated by an ELISA in a TC-CM or in the control medium. Results were expressed as mean ± SEM of seven independent experiments; ****p < 0.001; ***p < 0.005. D-F. Neutrophil survival in a TPC1-derived (D-E) or 8505c-derived (F-G) conditioned medium was evaluated in the presence of an anti-GM-CSF blocking antibody or the relative isotype control (10 μg/ml). At 24 hours, live cells were stained with FITC-conjugated annexin V and PI and analyzed by flow cytometry. Figs E and G illustrate representative flow cytometric panels of one out of five independent experiments. The results were expressed as mean ± SEM of five independent experiments; **p < 0.01; *p < 0.05. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/29953504), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human GM‑CSF Antibody

Application
Recommended Usage

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed human peripheral blood mononuclear cells

Western Blot

1 µg/mL
Sample: Recombinant Human GM-CSF (Catalog # 215-GM)
under non-reducing conditions only

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 0.075-0.45 µg/mL in the presence of 0.1 ng/mL Recombinant Human GM‑CSF.

Reviewed Applications

Read 2 reviews rated 4.5 using MAB215 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.

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

Granulocyte Macrophage Colony Stimulating Factor (GM-CSF) is a growth factor produced by a variety of lymphoid cells. It binds to receptor heterodimers consisting of a GM-CSF-specific alpha chain and the common beta chain that is shared by the high-affinity receptors for IL-3 and IL-5.

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

P04141 (Human)

Additional GM-CSF Products

Product Documents for Human GM‑CSF Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human GM‑CSF Antibody

For research use only

Citations for Human GM‑CSF Antibody

Customer Reviews for Human GM‑CSF Antibody (2)

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  • Human GM-CSF Antibody
    Name: Laboratorio Inmuno-Oncología
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: Melanoma tissue
    Species: Human
    Verified Customer | Posted 11/23/2020
    Human GM‑CSF Antibody MAB215
  • Human GM-CSF Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Pancreatic cancer cells
    Species: Human
    Verified Customer | Posted 05/02/2018

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Protocols

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