Mouse TNF-alpha Fluorescein-conjugated Antibody

Catalog # Availability Size / Price Qty
IC410F
Detection of TNF-alpha in Mouse Splenocytes by Flow Cytometry.
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Product Details
Citations (2)
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Mouse TNF-alpha Fluorescein-conjugated Antibody Summary

Species Reactivity
Mouse
Specificity
Detects mouse TNF-alpha.
Source
Monoclonal Rat IgG1 Clone # MP6-XT22
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
Mouse TNF-alpha
Formulation
Supplied in a saline solution containing BSA and Sodium Azide.
Label
Fluorescein (Excitation= 488 nm, Emission= 515-545 nm)

Applications

Recommended Concentration
Sample
Intracellular Staining by Flow Cytometry
10 µL/106 cells
See below

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

Intracellular Staining by Flow Cytometry View Larger

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 CFS-conjugated Monoclonal Antibody (Catalog # IC410F) and Rat anti-Mouse CD3 APC-conjugated Monoclonal Antibody (FAB4841A). Quadrant markers were set based on isotype control antibody (IC005F). 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.

Intracellular Staining by Flow Cytometry View Larger

Detection of TNF-alpha in Raw264 Mouse Cell Line by Flow Cytometry. Raw264 mouse macrophage cell line treated with 1 μg/mL LPS overnight was stained with Rat Anti-Mouse TNF-alpha Fluorescein-conjugated Monoclonal Antibody (Catalog # IC410F, filled histogram) or isotype control antibody (IC005F, open histogram). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3/Transcription Factor Fixation & Perm Kit (FC012). Staining was performed using our Staining Intracellular Molecules protocol.

Intracellular Staining by Flow Cytometry Detection of TNF-alpha antibody in EL‑4 Mouse Cell Line antibody by Flow Cytometry. View Larger

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 were stained with Rat Anti-Mouse TNF-alpha Fluorescein-conjugated Monoclonal Antibody (Catalog # IC410F, filled histogram) or isotype control antibody (Catalog # IC005F, open histogram). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.

Preparation and Storage

Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Protect from light. Do not freeze.
  • 12 months from date of receipt, 2 to 8 °C as supplied.

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.

Long Name
Tumor Necrosis Factor alpha
Entrez Gene IDs
7124 (Human); 21926 (Mouse); 24835 (Rat); 397086 (Porcine); 280943 (Bovine); 403922 (Canine); 102139631 (Cynomolgus Monkey); 100033834 (Equine); 493755 (Feline); 100009088 (Rabbit)
Alternate Names
APC1 protein; Cachectin; Cachetin; DIF; TNF; TNF, monocyte-derived; TNFA; TNF-A; TNFalpha; TNF-alpha; TNF-alphacachectin; TNFATNF, macrophage-derived; TNFG1F; TNFSF1A; TNFSF2; TNFSF2TNF superfamily, member 2; tumor necrosis factor (TNF superfamily, member 2); tumor necrosis factor alpha; Tumor necrosis factor ligand superfamily member 2; tumor necrosis factor; tumor necrosis factor-alpha

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Citations for Mouse TNF-alpha Fluorescein-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.

2 Citations: Showing 1 - 2
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  1. Transgenic Overexpression of Galectin-3 in Pancreatic beta Cells Attenuates Hyperglycemia in Mice: Synergistic Antidiabetic Effect With Exogenous IL-33
    Authors: N Jovicic, I Petrovic, N Pejnovic, B Ljujic, M Miletic Ko, S Pavlovic, I Jeftic, A Djukic, I Srejovic, V Jakovljevi, ML Lukic
    Frontiers in Pharmacology, 2021-11-05;12(0):714683.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  2. Tumor necrosis factor-alpha does not modulate ischemia/reperfusion injury in naïve myocardium but is essential for the development of late preconditioning.
    Authors: Dawn B, Guo Y, Rezazadeh A, Wang OL, Stein AB, Hunt G, Varma J, Xuan YT, Wu WJ, Tan W, Zhu X, Bolli R
    J. Mol. Cell. Cardiol., 2004-07-01;37(1):51-61.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC-P

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