Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Intracellular Staining by Flow Cytometry

Cited:

Flow Cytometry, Immunocytochemistry

Label

Phycoerythrin (Excitation = 488 nm, Emission = 565-605 nm)

Antibody Source

Monoclonal Mouse IgG1 Clone # 6402
View all TNF-alpha Primary Antibodies »

Product Specifications

Immunogen

Recombinant human TNF-alpha

Specificity

Detects cytoplasmic forms of human TNF-alpha.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Scientific Data Images for Human TNF‑ alpha PE‑conjugated Antibody

Detection of TNF- alpha antibody in Human PBMCs antibody by Flow Cytometry.

Detection of TNF‑ alpha in Human PBMCs by Flow Cytometry.

Human peripheral blood mononuclear cells (PBMCs) treated with PMA and Ca2+ionomycin for 3.5 hours were stained with (A) Mouse Anti-Human TNF-alpha PE-conjugated Monoclonal Antibody (Catalog # IC210P) or (B) isotype control antibody (IC002P) and Mouse anti-Human CD3 APC-conjugated Monoclonal Antibody (FAB100A). 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 Staining Intracellular Molecules protocol.

Applications for Human TNF‑ alpha PE‑conjugated Antibody

Application
Recommended Usage

Intracellular Staining by Flow Cytometry

10 µL/106 cells
Sample: Human peripheral blood mononuclear cells (PBMCs) 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).

Spectra Viewer

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Flow Cytometry

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

Store the unopened product at 2 - 8° C. Do not use past expiration date. Protect from light.

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- is produced by several lymphoid cells as well as by astrocytes, endothelial cells, and smooth muscle cells. Human TNF-alpha consisits of a 35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane segment, and a 177 aa extracellular domain (ECD). Within the ECD, human TNF-alpha shares 97% aa sequence identity with rhesus and 71%-92% with bovine, canine, cotton rat, equine, feline, mouse, porcine, and rat 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 Human TNF‑ alpha PE‑conjugated Antibody

Certificate of Analysis

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Product Specific Notices for Human TNF‑ alpha PE‑conjugated Antibody

For research use only

Citations for Human TNF‑ alpha PE‑conjugated Antibody

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FAQs for Human TNF‑ alpha PE‑conjugated Antibody

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  • Q: Why do FAB210 and IC210 give different intracellular staining results?

    A: FAB210 was developed to detect cells that are expressing cell surface TNF-alpha and the antibody is able to recognize cells that have bound TNF-alpha on their surface through their receptor and membrane bound forms. The IC210 on the other hand detects the cytoplasmic form of TNF-alpha, that is cells which are actively secreting TNF-alpha intracellularly. If FAB210 is used for intracellular staining, there is a chance that antibody will report additive staining of cell surface TNF-alpha as well as intracellular TNF-alpha upon permeabilization.

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