Colony Stimulating Factor 2 (Granulocyte-Macrophage) (CSF2) antibody

Details for Product No. ABIN967465
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Antigen
Synonyms CSF2, GM-CSF, GMCSF, CSF, Csfgm, Gm-CSf, MGI-IGM, Gm-csf, Gmcsf
Reactivity
Human
(201), (113), (30), (20), (12), (12), (6)
Host
Rat
(132), (92), (86), (8), (2), (2)
Clonality (Clone)
Monoclonal ()
Conjugate
Un-conjugated
(31), (8), (8), (5), (3), (2), (2), (2), (2), (2), (2), (2), (1), (1)
Application
Flow Cytometry (FACS), Immunoprecipitation (IP), Western Blotting (WB)
(212), (179), (41), (38), (32), (31), (28), (25), (24), (20), (16), (10), (9), (6), (6), (4), (4), (4), (4), (3), (2), (1), (1), (1), (1)
Pubmed 6 references available
Quantity 0.1 mg
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Catalog No. ABIN967465
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Immunogen Recombinant human GM-CSF
Clone BVD2-21C11
Isotype IgG2a
Characteristics 1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to us for technical protocols.
3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
4. Ficoll-Paque is a trademark of Amersham Biosciences Limited.
5. Cy is a trademark of Amersham Biosciences Limited.
Purification Purified from tissue culture supernatant or ascites by affinity chromatography.
Alternative Name GM-CSF
Background The BVD2-21C11 monoclonal antibody specifically binds to human Granulocyte/Macrophage - Colony Stimulating Factor (GM-CSF). Human GM-CSF is encoded by the CSF2 gene and is also known as Colony Stimulating Factor 2. GM-CSF is produced by activated T lymphocytes, macrophages, endothelial cells, fibroblasts, stromal cells and other cell types including B lymphocytes, mast cells, eosinophils, and osteoblasts. GM-CSF stimulates the survival, proliferation and/or differentiation of various cell types including neutrophils, eosinophils, macrophages, dendritic cells, megakaryocytes, erythroid cells, endothelial cells and their precursors. The immunogen used to generate the BVD2-21C11 hybridoma was recombinant human GM-CSF. The BVD2-21C11 antibody has been reported to crossreact with GM-CSF from the rhesus monkey. BVD2-21C11 is a neutralizing antibody.
Synonyms: CSF2, Colony stimulating factor 2 (granulocyte-macrophage), CSF, GMCSF
Research Area Immunology, Innate Immunity, Cytokines
Application Notes Blocking Control for Intracellular Staining:
The purified BVD2-21C11 antibody (ABIN967465) can be used as a blocking control to demonstrate specificity of human GM-CSF staining by the PE-BVD2-21C11. To perform this control, the fixed/permeabilized cells (~ 1 million) can be incubated with 1 - 10 µg of unlabeled BVD2-21C11 antibody (ABIN967465) for 20 minutes at 4°C, prior to staining with PE-BVD2-21C11 antibody (eg, 0.1 - 0.5 µg mAb/ 1 million cells). The intracellular cytokine staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcalfe.

ELISA Detection:
The biotinylated BVD2-21C11 antibody, is useful as a detection antibody for a sandwich ELISA for measuring human GM-CSF protein levels. Biotinylated BVD2-21C11 antibody can be paired with the purified BVD2-23B6 antibody as the capture antibody, with recombinant human GM-CSF as the standard. This ELISA pair is recommended primarily for measuring cytokine from experimental cell culture systems.
Restrictions For Research Use only
Format Liquid
Concentration 0.5 mg/ml
Buffer Aqueous buffered solution.
Preservative Sodium azide
Precaution of Use This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Storage 4 °C
Supplier Images
anti-Colony Stimulating Factor 2 (Granulocyte-Macrophage) (CSF2) antibody Expression of GM-CSF by stimulated human peripheral blood mononuclear cells (PBMC). Ficoll™-separated human PBMC were stimulated with soluble anti-human CD3 antibody (1 µg/ml final concentration, UCHT1), recombinant human IL-2 (10 ng/ml final concentration) and recombinant human IL-4 (10 ng/ml final concentration) for 2 days. The cells were subsequently cultured in medium containing recombinant human IL-2 and recombinant human IL-4 for 3 days. Finally, the cells were harvested and stimulated for 6 hours with PMA (50 ng/ml final concentration, Sigma) and calcium ionophore A23187 (250 ng/ml final concentration, Sigma) in the presence of BD GolgiStop™ (2 µM final concentration). The cells were harvested, stained with PE-Cy™5 anti CD4, fixed, permeabilized, and subsequently stained with 0.25 µg of PE Rat anti-Human GM-CSF antibody (PE-BVD2-21C11, ABIN967465) by using the staining protocol (first panel). To demonstrate specificity of staining, the binding of the PE-BVD2-21C11 antibody was blocked by preincubation of the antibody conjugate with recombinant human GM-CSF (0.1 µg, Center panel) and by preincubation of the fixed/permeabilized cells with unlabeled BVD2-21C11 antibody (5 µg, ABIN967465, second panel) prior to staining. The quadrant markers for the bivariate dot plot were set based on the autofluorescence control and verified using the ligand-blocking and unlabeled antibody blocking controls.
anti-Colony Stimulating Factor 2 (Granulocyte-Macrophage) (CSF2) antibody (2) Preincubation of the antibody conjugate with recombinant human GM-CSF
anti-Colony Stimulating Factor 2 (Granulocyte-Macrophage) (CSF2) antibody (3) Preincubation of the fixed/permeabilized cells with unlabeled BVD2-21C11 antibody
Product cited in: Abrams: "Immunoenzymetric assay of mouse and human cytokines using NIP-labeled anti-cytokine antibodies." in: Current protocols in immunology / edited by John E. Coligan ... [et al.], Vol. Chapter 6, pp. Unit 6.20, 2008 (PubMed).

General Abrams, Roncarolo, Yssel et al.: "Strategies of anti-cytokine monoclonal antibody development: immunoassay of IL-10 and IL-5 in clinical samples." in: Immunological reviews, Vol. 127, Issue 9-10, pp. 5-24, 1992 (PubMed).

Kita, Ohnishi, Okubo et al.: "Granulocyte/macrophage colony-stimulating factor and interleukin 3 release from human peripheral blood eosinophils and neutrophils." in: The Journal of experimental medicine, Vol. 174, Issue 3, pp. 745-8, 1991 (PubMed).

Bacchetta, de Waal Malefijt, Yssel et al.: "Host-reactive CD4+ and CD8+ T cell clones isolated from a human chimera produce IL-5, IL-2, IFN-gamma and granulocyte/macrophage-colony-stimulating factor but not IL-4." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 144, Issue 3, pp. 902-8, 1990 (PubMed).

Masaki, Kano, Merrick et al.: "Studies on Paul-Bunnell (P-B) antigen-antibody system. II. P-B antigens in extracts of lymphoma-leukemia spleens and pathologic sera." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 127, Issue 1, pp. 5-8, 1981 (PubMed).

Prussin, Metcalfe: "Detection of intracytoplasmic cytokine using flow cytometry and directly conjugated anti-cytokine antibodies." in: Journal of immunological methods, Vol. 188, Issue 1, pp. 117-28, 1996 (PubMed).

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