Mouse TNF-alpha PE-conjugated Antibody Summary
Applications
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Scientific Data
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Detection of TNF-alpha in Mouse Splenocytes by Flow Cytometry. Mouse splenocytes (A) treated with PMA (50 ng/mL), Ca2+ Ionomycin (200 ng/mL) and Brefeldin A (5 μg/mL) for 4 hours or (B) untreated, were stained with Rat Anti-Mouse TNF-alpha PE-conjugated Monoclonal Antibody (Catalog # IC410P) and Rat anti-Mouse CD3 APC-conjugated Monoclonal Antibody (FAB4841A). Quadrant markers were set based on isotype control antibody (IC005P). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (FC005). Staining was performed using our protocol for Staining Intracellular Molecules.
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Detection of TNF‑ alpha in EL‑4 Mouse Cell Line by Flow Cytometry. EL-4 mouse lymphoblast cell line treated with anti-CD3/anti-CD28, PMA, and Calcium Ionomycin was stained with Rat Anti-Mouse TNF-a PE-conjugated Monoclonal Antibody (Catalog # IC410P, filled histogram) or isotype control antibody (Catalog # IC005P, open histogram). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (FC005). View our protocol for Staining Intracellular Molecules.
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Detection of TNF‑ alpha in Raw264.7 cells by Flow Cytometry. Raw264.7 cells were stained with Rat Anti-Mouse TNF‑ alpha PE‑conjugated Monoclonal Antibody (Catalog # IC410P, filled histogram) or isotype control antibody (Catalog # IC005P, open histogram). To facilitate intracellular staining, cells were fixed with FC012 and permeabilized with FoxP3 Perm. View our protocol for Staining Intracellular Molecules.
Preparation and Storage
Background: TNF-alpha
Tumor necrosis factor alpha (TNF-alpha, TNF- alpha, TNFA ), also known as Cachectin and TNFSF2, is the prototypic ligand of the TNF superfamily. It is a pleiotropic molecule that plays a central role in inflammation, immune system development, apoptosis, and lipid metabolism. TNF-alpha is produced by several lymphoid cells as well as by astrocytes, endothelial cells, and smooth muscle cells. Mouse TNF-alpha consists of a 35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane segment, and a 179 aa extracellular domain (ECD). Within the ECD, mouse TNF-alpha shares 94% aa sequence identity with rat and 70%-77% with bovine, canine, cotton rat, equine, feline, human, porcine, and rhesus TNF-alpha. TNF-alpha is produced by a wide variety of immune, epithelial, endothelial, and tumor cells. TNF-alpha is assembled intracellularly to form a noncovalently linked homotrimer which is expressed on the cell surface. Cell surface TNF-alpha can induce the lysis of neighboring tumor cells and virus infected cells, and it can generate its own downstream cell signaling following ligation by soluble TNFR I. Shedding of membrane bound TNF-alpha by TACE/ADAM17 releases the bioactive cytokine, a 55 kDa molecular weight soluble trimer of the TNF-alpha extracellular domain. TNF-alpha binds the ubiquitous 55-60 kDa TNF RI and the hematopoietic cell-restricted 80 kDa TNF RII, both of which are also expressed as homotrimers present on virtually all cell types. Both type I and type II receptors bind TNF-alpha with comparable affinity, although only TNF RI contains a cytoplasmic death domain which triggers the activation of apoptosis. Soluble forms of both types of receptors are released and can neutralize the biological activity of TNF-alpha.
Product Datasheets
Citations for Mouse TNF-alpha PE-conjugated Antibody
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Citations: Showing 1 - 2
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Hepatic recruitment of CD11b+Ly6C+ inflammatory monocytes promotes hepatic ischemia/reperfusion injury
Authors: Peng Song, Junbin Zhang, Yunwei Zhang, Zhiping Shu, Peng Xu, Long He et al.
International Journal of Molecular Medicine
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Myeloid dendritic cells (DCs) of mice susceptible to paracoccidioidomycosis suppress T cell responses whereas myeloid and plasmacytoid DCs from resistant mice induce effector and regulatory T cells.
Authors: Pina A, de Araujo E, Felonato M, Loures F, Feriotti C, Bernardino S, Barbuto J, Calich V
Infect Immun, 2013-01-22;81(4):1064-77.
Species: Mouse
Sample Types: Whole Cells
Applications: Flow Cytometry
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