Human GM-CSF Antibody (3209)

R&D Systems | Catalog # MAB215

R&D Systems

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 (3209)

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.
Human GM-CSF ELISA Standard Curve

Human GM-CSF ELISA Standard Curve

Recombinant Human GM‑CSF (Catalog # 215-GM) was serially diluted and captured by Mouse Anti-Human GM‑CSF Monoclonal Antibody (Catalog # MAB615) coated on a Clear Polystyrene Microplate (Catalog # DY990). Mouse Anti-Human GM‑CSF Monoclonal Antibody (Catalog # MAB215) 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 Human GM-CSF Antibody (3209)

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 (3209)

Certificate of Analysis

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Product Specific Notices for Human GM-CSF Antibody (3209)

For research use only

Citations for Human GM-CSF Antibody (3209)

Customer Reviews for Human GM-CSF Antibody (3209) (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 (3209) 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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