Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Mouse

Cited:

Mouse

Applications

Validated:

Intracellular Staining by Flow Cytometry

Cited:

Flow Cytometry, Immunohistochemistry-Paraffin

Label

Fluorescein (Excitation = 488 nm, Emission = 515-545 nm)

Antibody Source

Monoclonal Rat IgG1 Clone # MP6-XT22

View all TNF-alpha Antibodies »

Product Specifications

Immunogen

Mouse TNF-alpha

Specificity

Detects mouse TNF-alpha.

Clonality

Monoclonal

Host

Rat

Isotype

IgG1

Scientific Data Images for Mouse TNF-alpha Fluorescein-conjugated Antibody

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.

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.
Detection of TNF-alpha antibody in EL-4 Mouse Cell Line antibody by Flow Cytometry.

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.

Applications for Mouse TNF-alpha Fluorescein-conjugated Antibody

Application
Recommended Usage

Intracellular Staining by Flow Cytometry

10 µL/106 cells
Sample: EL‑4 mouse lymphoblast cell line treated with anti-CD3/anti-CD28, PMA, and Calcium Ionomycin and mouse splenocytes treated with PMA and Ca2+ ionomycin were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005) and Raw264  mouse macrophage cell line treated with LPS, fixed and permeabilized with FlowX FoxP3/Transcription Factor Fixation & Perm Kit (Catalog # FC012).

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Formulation

Supplied in a saline solution containing BSA and Sodium Azide.

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

Alternate Names

Cachetin, DIF, TNF, TNF-A, TNFA, TNFalpha, TNFG1F, TNFSF1A, TNFSF2

Entrez Gene IDs

7124 (Human); 21926 (Mouse); 24835 (Rat); 397086 (Porcine); 280943 (Bovine); 403922 (Canine); 102139631 (Cynomolgus Monkey); 100033834 (Equine); 493755 (Feline); 100009088 (Rabbit)

Gene Symbol

TNF

Additional TNF-alpha Products

Product Documents for Mouse TNF-alpha Fluorescein-conjugated Antibody

Product Datasheets

Certificate of Analysis

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Product Specific Notices for Mouse TNF-alpha Fluorescein-conjugated Antibody

For research use only

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